LC3B Antibody - BSA Free

Images

 
Genetic Strategies: Lysates of HeLa parental cell line and LC3B knockout HeLa cell line (KO) untreated (-) or treated (+) with 50 uM CQ for 18 hours. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ...read more
Genetic Strategies: LC3B was detected in immersion fixed CQ treated HeLa cells (left) but was not detected in LC3B knockout HeLa cells (right) using rabbit anti-human LC3B polyclonal antibody (Catalog ...read more
Biological Strategies: Contribution of autophagy to lipid droplet formation in WT and KO NSC34 cells.(A) WT and KO NSC 34 cells were treated with (+) or without (-) bafilomycin A (bafA) under basal or HBSS ...read more
TMEM16A inhibited AngII-induced autophagy in aortas. (D) Representative immunofluorescence staining of LC3 (green) and a-SMA (red) in aortas. Scale bars, 20 um. Quantification of LC3B puncta. Five random fields (2,500 ...read more
LC3 accumulation in muscle fibers from patients with PAD (row B - E). Normal fibers from a Non-PAD sample (row A). Scale bar = 100 uM. Image from verified customer review.
Localization of LC3 by immunohistochemical method. The immunohistochemical staining shows that LC3, CRF, and HIF-1alpha have an overlapping localization in villous and extravillous trophoblast. d: decidua; v: villi; ...read more
Detection of LC3B in mouse ES cell lysates using Rabbit anti- [Catalog # NB100-2220]. LC3 detected at a molecular weight of approximately 17 kDa. Atg5-/- ES cells from Dr. Noboru Mizushima [Mizushima, N. et al. J. Cell ...read more
ORP1L and VAP-A form cholesterol-dependent ER-AV contact sites MelJuSo cells expressing GFP or GFP-tagged ORP1L mutants were fixed and stained for LC3 and Calnexin. Y477 and D478 in the FFAT motif were mutated to ...read more
Biological Strategies: Lysates of mouse NIH3T3 and rat PC-12 cell lines untreated (-) or treated (+) with CQ. PVDF (Polyvinylidene difluoride) membrane was probed with 0.5 ug/mL rabbit anti-LC3B polyclonal ...read more
Biological Strategies: Detection of LC3I and LC3II in mouse cochlea cell line SV-K1 using Rabbit anti- [Catalog # NB100-2220]. Cells were treated with CQ (1 uM), and As2O3 (1 uM) for 24 hrs. Image from verified ...read more
Autophagy Investigation. (A) Representative WBs (Western Blots) of LC3B-I, LC3B-II and GAPDH loading control in DANs derived from 4 iPD patients and 4 healthy controls. (B) WBs from three independent experiments were ...read more
Biological Strategies: Expression of autophagic protein LC3 in BME-UV1 cells forming acinar structures on Matrigel. Representative images of Western blot analysis of LC3 in bovine MECs grown in 3D culture for 3, ...read more
GNA triggers the formation of autophagic markers in A549 and HeLa cell. Effects of GNA on LC3 protein. A549 cells were treated with various concentrations of GNA for 24 hours or 3 uM GNA for the indicated periods of ...read more
Autophagosomes accumulate in neurons of the VH gray matter at day 1 after SCI. Representative images of IHC staining for LC3 (green) and neuronal marker NeuN (red) in VH of gray matter from sham and SCI animals. ...read more
Autophagosomes accumulate in microglia and oligodendrocytes of the dorsal white matter adjacent to injury after SCI. Representative images of IHC staining for LC3 (green) and activated microglia marker CD11B (red) in ...read more
Staining of rat brain tissue section. Analysis using Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Image from verified customer review.
Human ovarian Cancer tissue stained using heat mediated antigen retrieval in pH 6.0 citrate buffer at 1:200 dilution. Image provided by verified customer review.
FFPE (Formalin-Fixed Paraffin-Embedded) tissue section of mouse brain using 1:200 dilution of Rabbit anti-LC3B antibody [Catalog # NB100-2220]. The specific signal of LC3 was detected using HRP-conjugated secondary ...read more
Analysis using the Biotin conjugate of Rabbit anti-LC3B Antibody [Catalog # NB100-2220]. Staining of brain, cerebral cortex, neurons with cell processes.
SCI leads to lysosomal dysfunction. Representative high magnification images ( x 60) of LC3 (green) and CTSD (red) staining in VH of gray matter from sham and day 1 SCI animals. Decrease in number and intensity of CTSD ...read more
Image shows a specific band for LC3B at a molecular weight of approximately 15 kDa in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
LILRB2 antagonism effects on the bactericidal function of the MDSCs are independent of autophagy. (B) Lysates of cells from panel A were collected at 18 h and analyzed using Western blot for the autophagy marker, ...read more
CQ-induced autophagy inhibition leads to Nrf2 pathway activation in C2C12 muscle cells. a. Treatment of differentiated C2C12 myotubes with autophagy inhibitor CQ increased the sarcoplasmic level of LC3-II and SQSTM1 ...read more
Accumulation of an autophagosome marker in the presynaptic terminals of NMJs of SOD1G93A mice (A) Representative confocal micrographs of immunohistochemistry showing accumulation of the autophagosome marker LC3 ...read more
CAIX expression in PDAC and association with outcomeA. Representative images of immunohistochemical staining showing: negative expression of CAIX in both tumor cell and stromal compartments (top left), positive ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Al, Av, Ba, Bv, Ca, Ch, ChHa, Gp, Ha, In, Pm, Pm, Rb, SyHa, ZeSpecies Glossary
Applications WB, Simple Western, ELISA, Flow, Func, ICC/IF, IHC, IP, PAGE, WB, ChIP, KD, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Format
BSA Free
Concentration
1.0 mg/ml

Order Details

LC3B Antibody - BSA Free Summary

Immunogen
Polyclonal LC3B Antibody was made to a synthetic peptide made to an N-terminal portion of the human LC3B protein sequence (between residues 1-100). [UniProt# Q9GZQ8]
Localization
Type I form of LC3B is cytoplasmic, whereas the type II form of LC3B binds to the autophagic membranes.
Marker
Autophagosome Marker
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
MAP1LC3B
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Chromatin Immunoprecipitation (ChIP)
  • ELISA
  • Flow Cytometry
  • Immunoblotting
  • Immunocytochemistry/ Immunofluorescence 1:200
  • Immunohistochemistry 1:200 - 1:400
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200 - 1:400
  • Immunoprecipitation 20 ug/500 ug of protein
  • Knockdown Validated
  • Knockout Validated
  • Proximity Ligation Assay
  • SDS-Page
  • Simple Western 1:50
  • Western Blot 0.5 - 2.0 ug/mL
Application Notes
Use in SDS-PAGE reported in scientific literature (PMID:34315875).
Theoretical MW
14.688 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control
Human Brain Cerebral Cortex Whole Tissue Lysate (Adult Whole Normal)
Human Brain Whole Tissue Lysate (Adult Whole Normal)
Neuro2a Chloroquine Treated / Untreated Cell Lysate
HeLa Chloroquine Treated / Untreated Cell Lysate
Control Peptide
LC3B Protein (NB100-2220PEP)
Reviewed Applications
Read 51 Reviews rated 4.6
using
NB100-2220 in the following applications:

Publications
Read Publications using
NB100-2220 in the following applications:

Reactivity Notes

Use in Rat reported in scientific literature (PMID:34622072). Use in Chinese Hamster reported in scientific literature (PMID:34332287). Mouse reactivity reported in scientific literature (PMID:32814898). Bovine reactivity reported in scientific literature (PMID: 24895572). Primate reactivity reported in scientific literature (PMID: 25142602). Canine reactivity reported in scientific literature (PMID: 25839646). Avian reactivity reported in scientific literature (PMID: 29546310). Hamster reactivity reported in scientific literature (PMID: 26423766). Rabbit reactivity reported in scientific literature (PMID: 26497211). The mouse detection has been reported to be weaker than the human. Immunogen displays the following percentage of sequence identity for non-tested species: Xenopus 84%. Invertebrate reactivity reported in scientific literature (PMID: 26716072 ). Monkey reactivity reported in scientific literature (PMID: 30324853). Guinea pig reactivity reported from a verified customer review. Rat reactivity reported in scientific literature (PMID: 30744518). Bacteria reactivity reported in scientific literature (PMID: 28783414). Chicken reactivity reported in scientific literature (PMID: 30649814). Porcine reactivity reported in scientific literature (PMID: 30789643). Golden Syrian hamster reactivity reported in scientific literature (PMID: 23180219). Zebrafish reactivity reported in scientific literature (PMID: 29185873). Human reactivity reported in scientific literature (PMID:33086573). Use in Alligator reported in scientific literature (PMID:32061056).

Packaging, Storage & Formulations

Storage
Store at -20C.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for LC3B Antibody - BSA Free

  • anti-LC3B
  • Apg8b
  • atg8f antibody
  • ATG8F
  • Autophagy-related protein LC3 B
  • Autophagy-related ubiquitin-like modifier LC3 B
  • lc3b autophagy marker
  • LC3B ihc
  • LC3B immunohistochemistry
  • LC3B immunoprecipitation
  • LC3B western blot
  • LC3B
  • lc3-i, ii
  • LC3II
  • MAP1 light chain 3-like protein 2
  • MAP1A/1BLC3
  • MAP1A/MAP1B LC3 B
  • MAP1LC3B
  • MAP1LC3B-a
  • microtubule associated protein 1 light chain 3 beta
  • microtubule associated protein 3 b
  • microtubule associated protein 3 beta
  • microtubule-associated proteins 1A/1B light chain 3
  • microtubule-associated proteins 1A/1B light chain 3B

Background

Autophagy (macroautophagy) is a catabolic process which targets intracellular components such as proteins and organelles for degradation. Originally described as a bulk degradation process, current research supports its selective nature (1). Selective autophagy targets specific cellular components for degradation including the endoplasmic reticulum (2) (ER-phagy), mitochondria (3) (mitophagy), peroxisomes (3) (pexophagy), ribosomes (4) (ribophagy) and bacteria (5) (xenophagy). Autophagy relies on a newly formed phagophore, a membrane structure which elongates, sequesters cellular content, and fuses to form a double membrane vesicle known as the autophagosome. Fusion of autophagosomes with lysosomes gives rise to the autophagolysosome, where cellular components are degraded by lysosome hydrolases (1).

Autophagic flux is supported by autophagy-related proteins (Atgs) initially identified in yeast (6,7). The core autophagy machinery is comprised of 17 Atg proteins that play specific roles in autophagosome formation. Among these Atg proteins, Atg8 is not only involved in autophagosome formation but also functions in cargo selection. In mammals, several Atg8 homologues have been identified including microtubule-associated protein 1 light chain 3 alpha, beta and gamma - LC3A, LC3B, and LC3C (8) respectively, as well as GABA type A receptor-associated protein (GABARAP), GABARAP-Like1, and GABARAP-Like2 (9). LC3 (predicted molecular weight 14kD) is ubiquitously expressed and undergoes posttranslational processing after synthesis. First, the cysteine protease Atg4 cleaves a carboxy terminal sequence to generate the cytosolic form LC3-I. Next, E1-like (Atg7) and E2-like (Atg3) enzymes conjugate phosphatidylethanolamine to the newly exposed carboxyterminal glycine, generating LC3-II. Finally, the Atg12-Atg5-Atg16L1 complex participates in LC3 lipidation and autophagosome formation (10). LC3B-I to LC3B-II conversion correlates with autophagosome number and is considered the best marker to monitor autophagy.

References

1. Yu, L., Chen, Y., & Tooze, S. A. (2018). Autophagy pathway: Cellular and molecular mechanisms. Autophagy. https://doi.org/10.1080/15548627.2017.1378838

2. Forrester, A., De Leonibus, C., Grumati, P., Fasana, E., Piemontese, M., Staiano, L., ... Settembre, C. (2019). A selective ER -phagy exerts procollagen quality control via a Calnexin- FAM 134B complex. The EMBO Journal. https://doi.org/10.15252/embj.201899847

3. He, X., Zhu, Y., Zhang, Y., Geng, Y., Gong, J., Geng, J., ... Zhong, H. (2019). RNF34 functions in immunity and selective mitophagy by targeting MAVS for autophagic degradation. The EMBO Journal. https://doi.org/10.15252/embj.2018100978

4. Mathai, B., Meijer, A., & Simonsen, A. (2017). Studying Autophagy in Zebrafish. Cells. https://doi.org/10.3390/cells6030021

5. Losier, T. T., Akuma, M., McKee-Muir, O. C., LeBlond, N. D., Suk, Y., Alsaadi, R. M., ... Russell, R. C. (2019). AMPK Promotes Xenophagy through Priming of Autophagic Kinases upon Detection of Bacterial Outer Membrane Vesicles. Cell Reports. https://doi.org/10.1016/j.celrep.2019.01.062

6. Nakatogawa, H., Suzuki, K., Kamada, Y., & Ohsumi, Y. (2009). Dynamics and diversity in autophagy mechanisms: Lessons from yeast. Nature Reviews Molecular Cell Biology. https://doi.org/10.1038/nrm2708

7. Tsukada, M., & Ohsumi, Y. (1993). Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae. FEBS Letters. https://doi.org/10.1016/0014-5793(93)80398-E

8. Wild, P., McEwan, D. G., & Dikic, I. (2014). The LC3 interactome at a glance. Journal of Cell Science. https://doi.org/10.1242/jcs.140426

9. Igloi, G. L. (2001). Cloning, expression patterns, and chromosome localization of three human and two mouse homologues of GABAA receptor-associated protein. Genomics. https://doi.org/10.1006/geno.2001.6555

10. Glick, D., Barth, S., & Macleod, K. F. (2010). Autophagy: Cellular and molecular mechanisms. Journal of Pathology. https://doi.org/10.1002/path.2697

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for LC3B Antibody (NB100-2220)(1669)

We have publications tested in 20 confirmed species: Human, Mouse, Rat, Alligator, Avian, Bacteria, Bovine, C. elegans, Canine, Chicken, Common marmoset, Drosophila melanogaster, Golden Syrian Hamster, Guinea Pig, Hamster, Invertebrate, Monkey, Porcine, Primate, Zebrafish.

We have publications tested in 23 applications: B/N, Chemotaxis, ELISA, EM, FLOW, Flow, Func, IB, ICC, ICC/IF, IF/IHC, IHC, IHC-Fr, IHC-P, IP, In Vivo, KD, KO, PLA, SDS-Page, SW, Simple Western, WB.


Filter By Application
B/N
(23)
Chemotaxis
(1)
ELISA
(2)
EM
(4)
FLOW
(11)
Flow
(1)
Func
(2)
IB
(8)
ICC
(1)
ICC/IF
(225)
IF/IHC
(65)
IHC
(16)
IHC-Fr
(15)
IHC-P
(42)
IP
(8)
In Vivo
(8)
KD
(4)
KO
(5)
PLA
(1)
SDS-Page
(1)
SW
(1)
Simple Western
(1)
WB
(1020)
All Applications
Filter By Species
Human
(645)
Mouse
(410)
Rat
(94)
Alligator
(1)
Avian
(1)
Bacteria
(3)
Bovine
(6)
C. elegans
(1)
Canine
(2)
Chicken
(6)
Common marmoset
(1)
Drosophila melanogaster
(1)
Golden Syrian Hamster
(1)
Guinea Pig
(2)
Hamster
(1)
Invertebrate
(1)
Monkey
(3)
Porcine
(10)
Primate
(2)
Zebrafish
(9)
All Species
Showing Publications 1 - 10 of 1669. Show All 1669 Publications.
Publications using NB100-2220 Applications Species
Lu Y. Mechanistic study on the inhibition of the late stage of basal autophagy progression by a natural compound (NAADP) and a synthetic small chemical (vacuolin-1). Thesis. 2014-01-01 (ICC/IF, WB, Human)

Details:
LC3B/MAP1LC3B antibody used in experiments involving HeLa cells for immunofluorescence analysis (ICC-IF, at 1:500 dilution) and Western blotting analysis (WB, at 1:1000 dilution),
ICC/IF, WB Human
Lee E, Tran S, Juliani J et al. BECLIN1 is essential for intestinal homeostasis Research Square 2023-11-08 [PMID: 38378743] (WB) WB
Figueroa K, Gross C, Atienza E et al. GGC expansion inZFHX3causes SCA4 and impairs autophagy medRxiv 2023-10-29 (WB) WB
Chedid J The role of autophagy in TLR2-mediated alpha-synuclein accumulation in neurons. Thesis 2023-01-01 (WB, ICC/IF, Human) WB, ICC/IF Human
Kim J, Bae J, Park N et al. Inhibition of ubiquitin specific peptidase 14 (USP14) promotes ER-phagy by inducing ER stress in human hepatoma HepG2 cells Animal Cells and Systems 2023-12-11 (WB) WB
Paparo L, Coppola S, Nocerino R et al. How advanced glycation end products could facilitate the occurrence of food allergy The Journal of allergy and clinical immunology 2023-11-30 [PMID: 38042501] (WB, Human) WB Human
Trovò L, Vaucher A, Pan Y et al. In vitro assessment of dietary bioactives for TFEB activation as a possible target to support cognitive and emotional wellbeing Journal of Functional Foods 2023-12-01
Wu MY, Ge YJ, Wang EJ et al. Enhancement of efferocytosis through biased FPR2 signaling attenuates intestinal inflammation EMBO molecular medicine 2023-11-22 [PMID: 37994307]
Li B, Ming H, Qin S et al. HSPA8 Activates Wnt/?-Catenin Signaling to Facilitate BRAF V600E Colorectal Cancer Progression by CMA-Mediated CAV1 Degradation Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2023-11-16 [PMID: 37973552]
Ling JYM. Statins and Protein Prenylation in the Retina Thesis. 2017-01-01 (Human) Human
Show All 1669 Publications.

Reviews for LC3B Antibody (NB100-2220) (51) 4.651

Average Rating: 4.6
(Based on 51 reviews)
We have 51 reviews tested in 8 species: Human, Mouse, Rat, Guinea Pig, Human and Mouse, Insect and Mouse, Other, murine.

Reviews using NB100-2220:
Filter by Applications
Immunocytochemistry/Immunofluorescence
(1)
WB
(40)
ICC
(2)
Simple Western
(3)
IHC-Fr
(1)
IF
(1)
IHC
(1)
IHC-P
(1)
IP
(1)
All Applications
Filter by Species
Human
(21)
Mouse
(19)
Rat
(4)
Guinea Pig
(1)
Human and Mouse
(2)
Insect and Mouse
(1)
Other
(2)
murine
(1)
All Species
Images Ratings Applications Species Date Details
  3
reviewed by:
Verified Customer
Immunocytochemistry/Immunofluorescence Human 06/02/2021
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Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedLung Mesenchymal stem cells
SpeciesHuman
LotEU

Comments

CommentsCells were permeabilized with 0,2% triton, fixed with 4% PFA, Blocked with donkey serum, incubated for 1 h RT, washed with PBS-tween 20, incubated with secondary antibody donkey anti-rabbit Alexa-fluor 488, washed and stained with phalloidin texas-red and Hoechst 33342, mounted with anti-fade and photographed in confocal microscopy.
Western Blot LC3B NB100-2220
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4
reviewed by:
Hongxue Shi
WB Mouse 10/02/2020
View

Summary

ApplicationWestern Blot
Sample Testedmouse liver lysate
SpeciesMouse
LotEP
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 09/15/2020
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Summary

ApplicationWestern Blot
Sample TestedLiver tissue
SpeciesMouse
LotDC
Western Blot LC3B NB100-2220
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1
reviewed by:
Verified Customer
WB Insect and Mouse 09/10/2020
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Summary

ApplicationWestern Blot
Sample TestedC2C12 mouse myoblast cell line,Manduca sexta intersegmental muscle
SpeciesInsect and Mouse
LotEP

Comments

Comments20ug total protein was loaded on a 12% acrylamide gel. The blot was treated with 1:2000 dilution of antibody overnight at 4C, followed by HRP-labeled secondary and ECL detection.
*Low Rating Note: Novus Innovators Program - new species or application used on a primary antibody.
Western Blot LC3B NB100-2220
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4
reviewed by:
Verified Customer
WB Human 06/11/2019
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Summary

ApplicationWestern Blot
Sample Tested20 ug whole cell lysate
SpeciesHuman
LotCN

Comments

Comments1;1000 dilution of antibody in 20ug of protein worked well.
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Human 06/02/2019
View

Summary

ApplicationWestern Blot
Sample Tested20 ug whole cell lysate
SpeciesHuman
LotDC
  5
reviewed by:
Verified Customer
WB Human 03/04/2019
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Summary

ApplicationWestern Blot
Sample TestedTHP-1 whole cell lysate
SpeciesHuman
LotDQ-A1
Immunocytochemistry LC3B NB100-2220
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5
reviewed by:
Verified Customer
ICC Human 02/21/2019
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Summary

ApplicationImmunocytochemistry
Sample Testedhela cell
SpeciesHuman
LotEC
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 02/04/2019
View

Summary

ApplicationWestern Blot
Sample Testedmouse HSPCs
SpeciesMouse
LotAB-1
Western Blot LC3B NB100-2220
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4
reviewed by:
Verified Customer
WB Guinea Pig 02/04/2019
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Summary

ApplicationWestern Blot
Sample TestedGuinea pig ling, liver and heart
SpeciesGuinea Pig
LotDM-01
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 01/31/2019
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Summary

ApplicationWestern Blot
Sample Testedcochlea
SpeciesMouse
LotDJ2
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 01/29/2019
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Summary

ApplicationWestern Blot
Sample TestedHL-1 cell lysate
SpeciesMouse
LotDG-1
Simple Western LC3B NB100-2220
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5
reviewed by:
Marie Lassont
Simple Western Mouse 01/28/2019
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Summary

ApplicationSimple Western
Sample Testedmouse brain tisseu lysate
SpeciesMouse
LotDQ-1
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Human 07/17/2018
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Summary

ApplicationWestern Blot
Sample TestedTHP-1 whole cell lysate
SpeciesHuman
LotAQ-1

Comments

CommentsTHP-1 cells were treated with Rapamycin (100nm) for 24h. Total cell lysates were subjected to western blot. PVDF membrane were probed with 1mm/ml Human LC3B Antibody (NB100-2220) Antibody. A specific band was detected for LC3B I and LC3B II at approximately 13~15 kDa. This experiment was conducted under reducing conditions
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Human 06/07/2018
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Summary

ApplicationWestern Blot
Sample Testedfibroblasts whole cell extract,whole cell lysate
SpeciesHuman
LotDJ-1
Immunocytochemistry LC3B NB100-2220
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5
reviewed by:
Verified Customer
ICC Human 05/18/2018
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Summary

ApplicationImmunocytochemistry
Sample TestedHEK 293,hela cell
SpeciesHuman
LotDK-2
Western Blot LC3B NB100-2220
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5
reviewed by:
Dongkai Guo
WB Human and Mouse 05/10/2018
View

Summary

ApplicationWestern Blot
Sample TestedNIH3T3, HeLa, HEK293T, N2a neuroblastoma, Q7 striatal, Sy5Y neuroblastoma
SpeciesHuman and Mouse
LotDK-2
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 02/22/2018
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Summary

ApplicationWestern Blot
Sample Testedcochlea
SpeciesMouse
LotDQ-1
Western Blot LC3B NB100-2220
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4
reviewed by:
Verified Customer
WB Human and Mouse 11/25/2017
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Summary

ApplicationWestern Blot
Sample Testedmouse hepatocytes and HepG2 cells
SpeciesHuman and Mouse
LotDM-1
Immunohistochemistry-Frozen LC3B NB100-2220
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5
reviewed by:
Kate Kosmac
IHC-Fr Human 08/23/2017
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Summary

ApplicationImmunohistochemistry-Frozen
Sample TestedHuman skeletal muscle
SpeciesHuman
LotCX

Comments

CommentsFor LC3: Fresh-frozen sections were acetone fixed, washed with TBST(1% Tween), blocked for 1 hr with TBST, 5% NGS and incubated ON at 4C with primary antibody. LC3 staining was amplified with biotin followed by a directly conjugated streptavidin.
  5
reviewed by:
Verified Customer
WB Rat 07/14/2017
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Summary

ApplicationWestern Blot
Sample TestedNRK rat normal kidney cell line
SpeciesRat

Comments

CommentsUsed at 1:2500. Please see publication (Lee WK et al. Arch Toxicol 2017, in press) for details.

NOVUSCITED
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Human 04/12/2017
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Summary

ApplicationWestern Blot
Sample Tested20 ug whole cell lysate
SpeciesHuman
LotDH-2
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 03/23/2017
View

Summary

ApplicationWestern Blot
Sample TestedMacrophage
SpeciesMouse
LotDG-1
  4
reviewed by:
Paola Menichini
WB Human 03/14/2017
View

Summary

ApplicationWestern Blot
Sample TestedMDA MB 231 whole cell lysates and conditioned medium
SpeciesHuman
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB murine 02/14/2017
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Summary

ApplicationWestern Blot
Sample Testedmurine bone marrow derived macrophages
Speciesmurine
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Rat 01/18/2017
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Summary

ApplicationWestern Blot
Sample TestedChondrocyte cell lysate
SpeciesRat
LotCL
Immunofluorescence LC3B NB100-2220
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5
reviewed by:
Verified Customer
IF Mouse 01/29/2016
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Summary

ApplicationImmunofluorescence
Sample Testedmouse HSPCs
SpeciesMouse
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 01/29/2016
View

Summary

ApplicationWestern Blot
Sample Testedmouse HSPCs
SpeciesMouse
Simple Western LC3B NB100-2220
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5
reviewed by:
Verified Customer
Simple Western Mouse 01/21/2016
View

Summary

ApplicationSimple Western
Sample Testedmouse brain tisseu lysate
SpeciesMouse
Simple Western LC3B NB100-2220
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5
reviewed by:
Verified Customer
Simple Western Human 01/21/2016
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Summary

ApplicationSimple Western
Sample TestedSH-SY5Y cell lysate
SpeciesHuman
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Mouse 01/20/2016
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Summary

ApplicationWestern Blot
Sample TestedMouse eye tissue
SpeciesMouse
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Human 07/25/2015
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Summary

ApplicationWestern Blot
Sample TestedHuman prostate cancer cell line (PC3)
SpeciesHuman
Western Blot LC3B NB100-2220
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5
reviewed by:
Chunhui Sun
WB Human 01/30/2015
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Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesHuman
Western Blot LC3B NB100-2220
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4
reviewed by:
Verified Customer
WB Other 01/20/2015
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Summary

ApplicationWestern Blot
Sample TestedGuinea pig ling, liver and heart
SpeciesOther
Western Blot LC3B NB100-2220
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5
reviewed by:
Liyong Zhang
WB Mouse 06/26/2014
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Summary

ApplicationWestern Blot
SpeciesMouse
  4
reviewed by:
Lindsy Dickerson
WB Mouse 06/13/2014
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
Western Blot LC3B NB100-2220
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5
reviewed by:
Verified Customer
WB Human 03/27/2014
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Summary

ApplicationWestern Blot
SpeciesHuman
LotBK-1
Immunohistochemistry LC3B NB100-2220
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5
reviewed by:
Ashwini Katre
IHC Human 02/28/2014
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Summary

ApplicationImmunohistochemistry
Sample TestedOvarian cancer tissue
SpeciesHuman
LotBJ-1
Immunohistochemistry-Paraffin LC3B NB100-2220
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5
reviewed by:
Verified Customer
IHC-P Rat 01/16/2014
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedRat brain
SpeciesRat
Western Blot LC3B NB100-2220
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5
reviewed by:
Angela NIE
WB Mouse 02/12/2013
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Summary

ApplicationWestern Blot
Sample Testedmouse ventricle
SpeciesMouse
Western Blot LC3B NB100-2220
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5
reviewed by:
Myriam Elschami
WB Mouse 12/12/2012
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Summary

ApplicationWestern Blot
Sample TestedHL-1 cell lysate
SpeciesMouse
Western Blot LC3B NB100-2220
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4
reviewed by:
Angela NIE
WB Rat 01/09/2012
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Summary

ApplicationWestern Blot
Sample TestedH9C2 extract, Sample Amount: 20ug
SpeciesRat
LotA-K
  4
reviewed by:
Verified Customer
WB Human 12/05/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Mouse 11/21/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
  4
reviewed by:
Verified Customer
WB Human 11/21/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  5
reviewed by:
Verified Customer
WB Mouse 11/16/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
  5
reviewed by:
Verified Customer
WB Human 11/03/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Human 10/31/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  5
reviewed by:
Verified Customer
WB Human 10/31/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  4
reviewed by:
Devin Wolfe
WB Mouse 07/21/2010
View

Summary

ApplicationWestern Blot
Sample TestedMurine Blastocyst Derived Cells, Sample Amount: 30ug
SpeciesMouse
  5
reviewed by:
Verified Customer
IP Other 06/15/2009
View

Summary

ApplicationImmunoprecipitation
Sample TestedIP Sample
SpeciesOther

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3B Antibody (NB100-2220). (Showing 1 - 10 of 10 FAQs).

  1. Hello, somewhere on the LC3B antibody page I read it is recommended to use a 0.2uM membrane for western blot?
    • Yes, because LC3B-I is about 14-15 kDa and LC3B-II is even even smaller, the protein may slip through the 0.45 uM membrane and be lost during blotting. However, there has been success using 0.45 uM membranes; a 0.2 uM membrane is something to consider if no signal is detected.
  2. What are the best blocking conditions for NB100-2220?
    • The conditions we have had the most success with are 5% milk + 1% BSA as the block. Our lab also recommends doing an overnight incubation as this is how we run our QC tests in the lab at 4 degrees C.
  3. Is NB100-2220 the same or different from NBP-48614?
    • NB100-2220 is an anti-LC3 Ab. It is similar to NBP1-48614, however they are different Abs. NB100-2220 is specific for only LC3B where as NBP1-48614 is specific for only LC3A I would recommend NB100-2220 if you would like to detect LC3B as it has been in 48 published papers and it has very good reviews. NB100-2220 does detect both LCI and LCII.
  4. What proteins are recognized by this antibody? On the datasheet, the Specificity section states 'This antibody is specific for LC3B.' However, in the Gene Symbol is listed as MAP1LC3A and the protein sequence Q9H492 is for MAP1LC3A. Can you tell me which target the antibody recognizes?
    • This antibody was designed to detect both LC3I and LC3II, but we have experienced through our testing that it much more routinely picks up LC3B. It will sometimes pick up both bands, but it depends on how the cells are prepared often times if they are induced to express that second LC3A band, which is why we have it named as LC3B. We have other LC3 antibodies that do pick up both bands regularly.
  5. What is the isotype of this antibody? I would like to find an isotype control.
    • This is a polyclonal antibody from rabbit. The isotype of this antibody, and all rabbit antibodies, will be IgG.
  6. Does this antibody also recognizes LC3A and LC3C?
    • This antibody was raised using a synthetic peptide made to an N-terminal portion of the human LC3 protein sequence as immunogen and is expected to detect LC3B protein. The immunogenic sequence employed does not have any similarity to LC3A or LC3C and hence we do not expect any cross-reactivity. This product is one of our best sellers with great customer reviews, as well as with citations in at least 160 peer reviewed publications.
  7. Hello,I already have the antibody, but I was wondering if you could tell me the percent identity to a specific zebrafish LC3 protein. I know it says it works in zebrafish, but we have three and the one I am interested in is the one expressed maternally--- F1QKC7. Here is the Alignment to the human protein your antibody was made. Would you just look at it quick and tell me if the 10 aa peptide is one that is highly similar (like for example aa's 66-71). http://www.uniprot.org/align/2013102541KABHU792. LC3 Antibody (NB100-2220) It is very closely related to my maternal expressed protein. I was just wanting to confirm that it is the right thing and not background staining that I am seeing.
    • Based on the actual immunogen sequence, I am only seeing a 31% homology to the zebrafish sequence you have provided. While the homology is low, there is still a possibility that the antibody could recognize the zebrafish protein. Just for your information, the zebrafish sequence we used for validation was accession number Q7ZUD8. I can offer a few suggestions to verify specific staining in your samples: If it is possible, I would verify by western blot with your tissue. If you can validate by two applications, the staining seen is most likely specific. With the staining, I would run a secondary only or isotype control to ensure that any background staining is not coming from the secondary antibody or other interaction. If you do have access to a knock-down or knock-out, this would be the most ideal way to verify specific staining.
  8. We just bought the LC3 antibody (NB100-2220) from your company, and this product arrived at our lab today. Unfortunately, I just found that the label on the antibody vial showed "pAB LC3 antibody", so we are confused with the "p", what does "P"mean, usually it means"phosphorylated". So, I am wondering what does "p" mean, as the phosphorylated antibody is not we wanted. BTW, the datasheet of this product never mentioned the "phosphorylated"
    • I am sorry for any confusion. "pAB" stands for polyclonal antibody. Our NB100-2220 is not phosphorylated.
  9. I want to know how the lc3 antibody (nb100-2220) was produced and purified?
    • Of course. It is immunogen affinity purified. The immunogen is A synthetic peptide made to an N-terminal portion of the human LC3 protein sequence (between residues 1-100)
  10. Why are there two bands present when I use LC3B antibody NB100-2220 for a western blot of LC3B?
    • LC3 is a soluble protein with a molecular mass of ∼17 kDa and is distributed ubiquitously in eukaryotes. It is expressed as the splice variants LC3A, LC3B, and LC3C. All LC3 isoforms undergo post-translational modifications, especially PE conjugation (lipidation) during autophagy. Upon autophagic signal, the cytosolic form of LC3 (LC3-I) is conjugated to PE to form LC3-PE conjugate (LC3-II), which is recruited to the autophagosomal membranes. For more information, you can refer to our resource page

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Additional LC3B Products

Array NB100-2220

Research Areas for LC3B Antibody (NB100-2220)

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Blogs on LC3B. Showing 1-10 of 26 blog posts - Show all blog posts.

Autophagy and RAS signaling: Clinical implications
By Christina Towers, PhD The cellular recycling process known as autophagy is currently being targeted in over 60 clinical trials focused on treating different types of cancer1. To date, the only autophagy-targeted ...  Read full blog post.

Methamphetamine with HIV induces mitochondrial dysfunction and neuronal injury through oxidative stress
By Jamshed Arslan, Pharm. D., PhD. December 1 is the World AIDS Day. Despite the combination antiretroviral therapy, 10-25% of Human Immunodeficiency Virus (HIV)-positive individuals report neurocognitive impairm...  Read full blog post.

Autophagy and Metastasis
  By Christina Towers, PhD The majority of cancer patients die from metastatic disease at secondary sites. The threshold to undergo metastasis is high. Only a minority of cancer cells acquire invasive phenotypes...  Read full blog post.

Optogenetic Control of Mitophagy: AMBRA1 based mitophagy switch
By Christina Towers, PhD Mitophagy in the BrainSelective autophagic degradation of damaged mitochondria, known as mitophagy, has been described as a cyto-protective process. Accordingly, defects in mitophagy h...  Read full blog post.


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How to visualize autophagy by microscopy
By Christina Towers, PhD Autophagy is a recycling process that relies on the formation of a unique organelle termed an autophagosome. An elegant way to monitor autophagy is through various microscopy techniques to...  Read full blog post.


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Showing 1-10 of 26 blog posts - Show all blog posts.
mFluor Violet Conjugated Antibodies

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Recent Reviews

4.6
5
36
4
13
3
1
2
0
1
1

Verified Customer
06/02/2021
Application: Immunocytochemistry/Immunofluorescence
Species: Human

Hongxue Shi
10/02/2020
Application: WB
Species: Mouse

Verified Customer
09/15/2020
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol MAP1LC3B