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LC3A Antibody

 (19 Publications)
 (2 Reviews)
 
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Western Blot: LC3A Antibody [NBP1-19167] - Total protein from HeLa and Neuro2A cells treated with or without 50 uM chloroquine for 24 hours was separated on a 4-15% gel by SDS-PAGE, transferred to 0.2 um PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL anti-LC3A in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the detection LC3 II upon chloroquine treatment.
Biological Strategies: Western Blot: LC3A Antibody [NBP1-19167] - Total protein from HeLa and Neuro2A cells treated with or without 50 uM chloroquine for 24 hours was separated on a 4-15% gel by SDS-PAGE, ...read more
Immunocytochemistry/ Immunofluorescence: LC3A Antibody [NBP1-19167] - LC3/MAP1 [NBP1-19167] - LC3 antibody was tested in HeLa cells with Dylight 488 (green). Cells were treated overnight with 50 uM chloroquine to induce ...read more
Immunohistochemistry-Paraffin: LC3A Antibody [NBP1-19167] - Analysis of a FFPE tissue section of mouse brain using LC3 antibody at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB ...read more
Western Blot: LC3/MAP1LC3A Antibody [NBP1-19167] - Human brain lysate.
Immunohistochemistry-Paraffin: LC3A Antibody [NBP1-19167] - Analysis of a FFPE tissue section of mouse liver using LC3 antibody at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB ...read more
Simple Western: LC3A Antibody [NBP1-19167] - Image shows a specific band for LC3 in 0.5 mg/mL of Neuro2A lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Product Details

Summary
Reactivity Hu, Mu, Rt, Bv, ZeSpecies Glossary
Applications WB, Simple Western, Flow, ICC/IF, IHC
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml
Validated by:
   

Biological Strategies

      

Order Details

View Available Conjugates
Catalog# & Conjugate Size Price
Alexa Fluor 350
NBP1-19167AF350
Alexa Fluor 405
NBP1-19167AF405
Alexa Fluor 488
NBP1-19167AF488
Alexa Fluor 532
NBP1-19167AF532
Alexa Fluor 594
NBP1-19167AF594
Alexa Fluor 647
NBP1-19167AF647
Alexa Fluor 700
NBP1-19167AF700
Alexa Fluor 750
NBP1-19167AF750
Allophycocyanin
NBP1-19167APC
CoraFluor 1
NBP1-19167CL1
DyLight 350
NBP1-19167UV
DyLight 405
NBP1-19167V
DyLight 594
NBP1-19167DL594
DyLight 680
NBP1-19167FR
DyLight 755
NBP1-19167IR
Janelia Fluor 525
NBP1-19167JF525
Janelia Fluor 549
NBP1-19167JF549
Janelia Fluor 585
NBP1-19167JF585
Janelia Fluor 635
NBP1-19167JF635
Janelia Fluor 646
NBP1-19167JF646
Janelia Fluor 669
NBP1-19167JF669
PE
NBP1-19167PE
PerCP
NBP1-19167PCP
mFluor Violet 450 SE
NBP1-19167MFV450
mFluor Violet 500 SE
NBP1-19167MFV500
mFluor Violet 610 SE
NBP1-19167MFV610
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LC3A Antibody Summary

Immunogen
Partial recombinant protein made to an N-terminal portion of the human LC3A protein [Swiss-Prot# Q9H492].
Localization
Autophagosomes. Cytoplasm. Intracytoplasmic membrane; Lipid-anchor. Cytoplasmic vesicle autophagosome membrane; Lipid-anchor. Note= LC3-II binds to the autophagic membranes.
Marker
Autophagosome Marker
Specificity
Although specificity between this LC3A Antibody and LC3B has not been tested, this antibody was created to a peptide that has 100% identity to LC3A and 62% identity to LC3B.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
MAP1LC3A
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.
Learn about the Innovator's Reward

Applications/Dilutions

Dilutions
  • Flow Cytometry 1:100
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry 1:100 - 1:400
  • Immunohistochemistry-Frozen reported in scientific literature (PMID 23936035)
  • Immunohistochemistry-Paraffin 1:100 - 1:400
  • Simple Western 1:40
  • Western Blot 1:2500
Application Notes
In WB, bands are seen at approx. 14-17 kDa. In ICC/IF, autophagosome formation has been seen in HeLa cells after treatment with 50 uM chloroquine.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Theoretical MW
14 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 Whole Tissue Lysate (Adult Whole Normal)
Human Brain Thalamus Whole Tissue Lysate (Adult Whole Normal)
Neuro2a Chloroquine Treated / Untreated Cell Lysate
HeLa Chloroquine Treated / Untreated Cell Lysate
Reviewed Applications
Read 2 Reviews rated 4.5
using
NBP1-19167 in the following applications:
Publications
Read Publications using
NBP1-19167 in the following applications:

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for LC3A Antibody

  • Apg8
  • APG8a
  • Apg8p3
  • ATG8E
  • Autophagy-related protein LC3 A
  • Autophagy-related ubiquitin-like modifier LC3 A
  • LC3
  • LC3A
  • MAP1 light chain 3-like protein 1
  • MAP1A/1B light chain 3 A
  • MAP1A/MAP1B LC3 A
  • MAP1A/MAP1B light chain 3 A
  • MAP1ALC3
  • MAP1BLC3
  • MAP1LC3A
  • Microtubule-associated protein 1 light chain 3 alpha
  • microtubule-associated proteins 1A/1B light chain 3
  • microtubule-associated proteins 1A/1B light chain 3A
  • MLP3A

Background

Human Microtubule-associated Protein 1A/1B Light Chain 3A (MAP1LC3A), also called LC3A for short, is a 121 amino acid (aa) protein with a theoretic molecular weight of ~14 kDa. LC3A belongs to the LC3 subfamily of Autophagy-related 8 (Atg8) proteins, which also includes LC3B and LC3C (1). The process of autophagy is the bulk degradation of proteins and organelles. Whether these three proteins have distinct roles in autophagy remains unclear, but they do have unique subcellular expression (2). Specifically, LC3A shows perinuclear and nuclear localization (2). The Atg8 family members share a similar structure of two amino-terminal alpha-helices and a ubiquitin-like core but are unique in aa sequence (1). LC3 utilizes a ubiquitin-like conjugation system to covalently bind to phosphatidylethanolamine (PE), also called lipidation, which is mediated by a series of steps (1, 3). Briefly, unprocessed, cytosolic LC3 (pro-LC3) is cleaved by the cysteine protease Atg4 to expose a c-terminal glycine (Gly) residue (LC3-I); the Gly is activated by E1-like enzyme Atg7, transferred to E2-like enzyme Atg3, and an E3-like complex facilitates the conjugation of LC3 with PE (LC3-II), incorporating it into the phagophore membrane during autophagosome formation (1, 3). The recruitment of LC3 to the phagophore is thought to mediate membrane elongation and closure (1, 3). Additionally, LC3 plays a role in recruiting cargo (protein aggregates, pathogens, and organelles) to autophagosomes and delivery for lysosomal degradation (1).

The process of autophagy is associated with a variety of diseases including neurodegenerative diseases, neuromuscular, tumorigenesis, and viral and bacterial infections (4). LC3 is a useful marker of autophagy in both healthy and diseased cells (4). Interestingly, LC3A has two variants (v1 and v2) which differ in N-terminal sequence due to the varying transcriptional start sites (5). One particular study found that LC3Av1, but not v2 or LC3B, was silenced in various cancer cell lines due to aberrant DNA methylation and re-expression of LC3Av1 in LC3Av1-silenced cells inhibited tumor growth, where overall findings suggest a possible tumor-suppressive role (5).

Alternative names for LC3A include Apg8, APG8a, ATG8E, Autophagy-related protein LC3 A, Autophagy-related ubiquitin-like modifier LC3 A, MAP1A/1B light chain 3 A, microtubule-associated proteins 1A/1B light chain 3, and MLP3A.

References

1. Shpilka, T., Weidberg, H., Pietrokovski, S., & Elazar, Z. (2011). Atg8: an autophagy-related ubiquitin-like protein family. Genome biology. https://doi.org/10.1186/gb-2011-12-7-226

2. Koukourakis, M. I., Kalamida, D., Giatromanolaki, A., Zois, C. E., Sivridis, E., Pouliliou, S., Mitrakas, A., Gatter, K. C., & Harris, A. L. (2015). Autophagosome Proteins LC3A, LC3B and LC3C Have Distinct Subcellular Distribution Kinetics and Expression in Cancer Cell Lines. PloS one. https://doi.org/10.1371/journal.pone.0137675

3. Weidberg, H., Shvets, E., & Elazar, Z. (2011). Biogenesis and cargo selectivity of autophagosomes. Annual review of biochemistry. https://doi.org/10.1146/annurev-biochem-052709-094552

4. Tanida, I., Ueno, T., & Kominami, E. (2004). LC3 conjugation system in mammalian autophagy. The international journal of biochemistry & cell biology. https://doi.org/10.1016/j.biocel.2004.05.009

5. Schaaf, M. B., Keulers, T. G., Vooijs, M. A., & Rouschop, K. M. (2016). LC3/GABARAP family proteins: autophagy-(un)related functions. FASEB journal : official publication of the Federation of American Societies for Experimental Biology. https://doi.org/10.1096/fj.201600698R

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.

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Publications for LC3A Antibody (NBP1-19167)(19)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 5 applications: ICC/IF, IF/IHC, IHC-Fr, IHC-P, WB.


Filter By Application
ICC/IF
(2)
IF/IHC
(2)
IHC-Fr
(1)
IHC-P
(3)
WB
(10)
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Human
(10)
Mouse
(8)
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Showing Publications 1 - 10 of 19. Show All 19 Publications.
Publications using NBP1-19167 Applications Species
Korski KI, Kubli DA, Wang BJ et al. Hypoxia Prevents Mitochondrial Dysfunction and Senescence in Human c-Kit+ Cardiac Progenitor Cells. Stem Cells 2019-01-10 [PMID: 30629785] (ICC/IF, Human) ICC/IF Human
Jiang Z, Bo L, et al. Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy. Cell Death Dis 2018-08-28 [PMID: 30154452] (WB, Mouse) WB Mouse
Yang A, Herter-Sprie G, Zhang H, et al. Autophagy Sustains Pancreatic Cancer Growth through Both Cell-Autonomous and Nonautonomous Mechanisms. Cancer Discov. 2018-01-09 [PMID: 29317452] (IF/IHC, Mouse) IF/IHC Mouse
Kohler LJ, Reed SR, Sarraf SA et al. Effector Protein Cig2 Decreases Host Tolerance of Infection by Directing Constitutive Fusion of Autophagosomes with the Coxiella-Containing Vacuole MBio 2016 Jul 20 [PMID: 27435465] (WB). Elife. 2016-07-20 [PMID: 27435465] (WB)

Details:
This citation used the PE version of this antibody. 		WB
Saito T, Asai K, Sato S et al. Autophagic vacuoles in cardiomyocytes of dilated cardiomyopathy with initially decompensated heart failure predict improved prognosis Autophagy 2016-02-18 [PMID: 26890610] (IHC-P, Human) IHC-P Human
Campos T, Ziehe J, Palma M et al. Rheb promotes cancer cell survival through p27Kip1-dependent activation of autophagy Mol. Carcinog. 2015-01-15 [PMID: 25594310] (WB, Human) WB Human
Yang Annan, Rajeshkumar N V, Wang Xiaoxu et al. Autophagy is critical for pancreatic tumor growth and progression in tumors with p53 alterations. Cancer Discov. 2014-05-29 [PMID: 24875860]
Rao S, Tortola L, Perlot T et al. A dual role for autophagy in a murine model of lung cancer. Nat Commun. 2014-01-01 [PMID: 24445999] (IHC-P, Mouse) IHC-P Mouse
Giovannini M, Bonne NX, Vitte J et al. mTORC1 inhibition delays growth of neurofibromatosis type 2 schwannoma. Neuro-oncology 2014-01-10 [PMID: 24414536] (IHC-P, Mouse) IHC-P Mouse
Bruntz RC, Taylor HE, Lindsley CW, Brown HA. Phospholipase D2 mediates survival signaling through direct regulation of Akt in glioblastoma cells. J Biol Chem. 2013-11-20 [PMID: 24257753] (WB, Human) WB Human
Show All 19 Publications.

Reviews for LC3A Antibody (NBP1-19167) (2) 4.52

Average Rating: 4.5
(Based on 2 reviews)
We have 2 reviews tested in 2 species: Mouse, Rat.

Reviews using NBP1-19167:
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ICC
(1)
WB
(1)
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Mouse
(1)
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(1)
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Images Ratings Applications Species Date Details
Immunocytochemistry LC3A NBP1-19167
Enlarge 		4
reviewed by:
Tania Campos
 ICC Mouse 06/12/2015
View

Summary

ApplicationImmunocytochemistry
Sample TestedMEF cells
SpeciesMouse
LotC-1
Western Blot LC3A NBP1-19167
Enlarge 		5
reviewed by:
Verified Customer
 WB Rat 02/24/2015
View

Summary

ApplicationWestern Blot
Sample TestedRat bone marrow stromal cells
SpeciesRat

Product General Protocols

View specific protocols for LC3A Antibody (NBP1-19167): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for LC3A Antibody (NBP1-19167). (Showing 1 - 3 of 3 FAQs).

  1. May we ask if it is possible to perform IF to stain LC3-I and LC3-II separately with two different fluorescent colors?
    • Yes, it is possible to perform IF stain for LC3-I and LC3-II separately with two different fluorescent colors! You will have to use two different primary antibodies and in order to avoid any potential background/cross reactivity issues, I would suggest that you employ conjugated primary antibodies for the testing. 1. Our LC3I antibody (NBP1-78964) has been designed to specifically detect the cytosolic form of the LC3 protein which is actually LC3 I (Note: LC3-II binds to the autophagic membranes). 2. There is not even a single antibody to our knowledge that would exclusively detect the LC3 II form, and you would have to detect LC3II/ autophagic membranes form with an antibody which detects LC3 I/LC3II together. Therefore you may opt second antibody from one of the followings: LC3 Antibody (NB100-2220), LC3 Antibody (NB100-2331), LC3 Antibody (NBP1-19167). All of these mentioned catalog #s come with different options for their conjugated forms and you may select appropriate conjugated forms for performing the IF staining using our explained criteria.
  2. We have received an antibody against LC3 (# NBP1-19167) and I need a precision regarding its specificity. According to the datasheet, the antibody was raised against a peptide 100% identical to LC3A and 62% to LC3B. My question is: Is LC3A used as a synonym for LC3-I and LC3B as a synonym of LC3-II?
    • LC3A is not a synonym for LC3-I or LC3-II. When we refer to LC3-I and -II, we actually mean LC3B-I and LC3B-II. I hope this helps, but please let me know if you have any questions.
  3. We purchased an antibody against LC3 (NBP1-19167) from your company. According to the information found on your web site, three papers used this antibody to detect both the LC3-I and LC3-II form of the protein. The catalog number of the antibody is not mentioned in the papers. How can I be sure that it is the one used by the authors? It is important since I want to know if this antibody NBP1-19167 recognize both LC3 forms.
    • Our publication evaluation team is highly qualified and we found that these publications were cited under # NBP1-19167 after consulting the archived order information of the customers. As far as the detection of the two forms is concerned, any given LC3 antibody should detect LC3-I as well as LC3-II forms because the only difference between the two forms is that LC3-II carries the phosphatidylethanolamine (PE) conjugate whereas LC3-I does not have a PE conjugate. In nutshell, following translation, the unprocessed form of LC3 (pro-LC3) is proteolytically cleaved by Atg4 protease, resulting in the cytosolic form of LC3 (LC3-I) with a carboxyterminal exposed glycine. Upon induction of autophagy, the exposed glycine of LC3-I is conjugated by Atg7 (an E1-like activity), Atg3 (an E2-like conjugating activity) and by Atg12-Atg5-Atg16L multimers (E3-like ligase activity) to the highly lipophilic PE moiety to generate LC3-II (which is recruited to autophagosomal membranes). Therefore, the presence of both bands (LC3-I & LC3-II) will depend upon the extent of autophagy in your particular samples.

Control Lysate(s)

Human Brain Whole Tissue Lysate (Adult Whole Normal)
Human Brain Thalamus Whole Tissue Lysate (Adult Whole Normal)
Neuro2a Chloroquine Treated / Untreated Cell Lysate
HeLa Chloroquine Treated / Untreated Cell Lysate

Secondary Antibodies

 

Isotype Controls

Additional LC3A Products

Research Areas for LC3A Antibody (NBP1-19167)

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Blogs on LC3A.

Check out the latest blog posts on LC3A.

  Read full blog post.
Losing memory: Toxicity from mutant APP and amyloid beta explain the hippocampal neuronal damage in Alzheimer's disease
 By Jamshed Arslan Pharm.D.  Alzheimer's disease (AD) is an irreversible brain disorder that destroys memory and thinking skills. The telltale signs of AD brains are extracellular deposits of amy...  Read full blog post.
Nuclear LC3: Why is it there and what is it doing?
By Christina Towers, PhD. Cells use the complex process of autophagy to degrade and recycle cytoplasmic material.  There are over 20 proteins that have been implicated in this process and appropriately named core ...  Read full blog post.
Why LC3B Antibodies Make Ideal Autophagosomes Membrane Markers
The human form of microtubule-associated protein light chain 3 (LC3) is expressed as 3 splice variants LC3A, LC3B, and LC3C.1 LC3B is a subunit of the MAP1A and MAP1B microtubule-binding proteins and plays a central role in autophagosome membrane stru...  Read full blog post.
mFluor Violet Conjugated Antibodies

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  ICC/IF Protocol
  IHC-P Protocol
  Western Blot Protocol
  SDS
  PDF Datasheet
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Recent Reviews

4.5
5
1
4
1
3
0
2
0
1
0
Immunocytochemistry LC3A NBP1-19167
Enlarge
 Tania Campos
06/12/2015
Application: ICC
Species: Mouse
read this review
Western Blot LC3A NBP1-19167
Enlarge
 Verified Customer
02/24/2015
Application: WB
Species: Rat
read this review

Bioinformatics

Gene Symbol MAP1LC3A
Entrez
Uniprot