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GAPDH Antibody (1D4)

Images

 
Analysis of GAPDH in human nucleus pulposus cell culture (40ug/lane) using anti-GAPDH antibody. LI-COR Odyssey imaging system was used for detection. Western blot image submitted by a verified customer review.
Analysis of of HeLa cells stained with GAPDH antibody, dilution 1:100 (Green). DAPI staining of nuclear DNA (Blue). The GAPDH antibody produces strong cytoplasmic staining of healthy cells.
Analysis of cell line lysates probed with mouse mAb to GAPDH (theoretical molecular weight: 36 kDa), NB300-221, dilution 1:2,000: [1] protein standard, [2] HEK293, [3] HeLa, [4] SH-SY5Y, [5] COS1, [6] NIH-3T3, and [7] ...read more
Human eye cell lysate. Antibody used at 1:500. WB image submitted by a verified customer review.
Analysis of GAPDH in chicken pectoralis major using anti-GAPDH antibody. The primary antibody was used at a dilution of 1:4000, incubated for 1 hour at room temperature. Western blot image submitted by a verified ...read more
Application of the approach to other cell lines. The remainder of the protocol from Figure 2 was then applied using 100 ug/mL digitonin as the optimal cytosolic extract in each case. An aliquot of each extract was then ...read more
Endothelial responses provoked by Hb-lipid interactions are attenuated by sulfide donors. Confluence HAoECs were exposed to the obtained Hb samples for 8 hours and VCAM-1 expression was determined by Western blotting. ...read more
Regulation of cell migration-associated proteins by NCKU-21 in A549 and CL1-5 cells. Protein expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 were analyzed in A549 cell treated with NCKU-21 for 24 hr. * P 80% ...read more
Immunodepletions and the effects of RNases and various transcription inhibitors on chromosome condensation and cell cycle state.(A) Western blot for CAP-E, XPB, and Tubulin in Mock or CAP-E depleted extracts. Dilution ...read more
Activation of the ATM‐Chk2‐p53 pathway upon CDDP treatmentAConfocal images showing the basal region of organ of Corti cultures treated with either culture medium alone or medium containing 10 μM CDDP for 1 day ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Av, Bv, Ca, Ch, ChHa, Dr, Eq, Fe, Ha, Ma-Op, Pm, Rb, Sh, ZeSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC, IP, ChIP, KD, Single-Cell Western
Clone
1D4
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Order Details

GAPDH Antibody (1D4) Summary

Description
This GAPDH antibody (1D4) is primarily used as a loading control in western blots due to the near global expression of GAPDH which can be attributed to its vital role in the glycolytic metabolic process.
Immunogen
This GAPDH antibody was developed against full length purified porcine GAPDH [UniProt# P00355]
Marker
Cytosolic Marker
Isotype
IgM
Clonality
Monoclonal
Host
Mouse
Gene
GAPDH
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) 1:10-1:500
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Frozen
  • Immunoprecipitation
  • Knockdown Validated
  • Simple Western 1:25
  • Single Cell Western
  • Western Blot 1:1000
Theoretical MW
36 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
GAPDH Overexpression Lysate
Reviewed Applications
Read 19 Reviews rated 4.4
using
NB300-221 in the following applications:

Publications
Read Publications using
NB300-221 in the following applications:

Reactivity Notes

Likely to react with other mammals as well. Hamster, zebrafish and feline reactivity reported by customer reviews. Chicken, rabbit, and Chinese hamster reactivity reported in scientific literature (PMID: 24725420, 17545481, and 24928958 respectively). Drosophila reactivity reported in scientific literature (PMID: 32878003)

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
50% PBS, 50% glycerol
Preservative
0.035% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for GAPDH Antibody (1D4)

  • 1D4 GAPDH
  • aging-associated gene 9 protein
  • Cytoplasm Marker
  • EC 1.2.1
  • EC 1.2.1.12
  • EC 2.6.99.-
  • G3PD antibody
  • G3PD
  • G3PDH
  • GAPD
  • GAPDH
  • GAPDPeptidyl-cysteine S-nitrosylase GAPDH antibody
  • glyceraldehyde 3-phosphate dehydrogenase
  • glyceraldehyde-3-phosphate dehydrogenase
  • MGC102544
  • MGC102546
  • MGC103190
  • MGC103191
  • MGC105239
  • MGC127711
  • MGC88685
  • OCAS, p38
  • OCT1 coactivator in S phase
  • Peptidyl-cysteine S-nitrosylase GAPDH

Background

Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a ubiquitous enzyme involved in glycolysis, converting glyceraldehyde-3-phosphate into 1,3 diphosphoglycerate. This constitutively expressed, homotetramer protein can be found in the nucleus and cytoplasm, and the monomer has a theoretical molecular weight of 36 kDa. Known as a housekeeping gene, GAPDH routinely serves as a control for real-time PCR (RT-PCR) and as a loading control for Western Blots (1-3). In addition to its role in glycolysis, GAPDH has multiple cellular functions including membrane trafficking, transcription activation, apoptosis, DNA repair, and has been implicated in various pathologies such as cancer and neurodegenerative diseases including Alzheimer's and Huntington's (4,5).

References

1) Barber RD, Harmer DW, Coleman RA, Clark BJ. (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 21(3):389-95. PMID: 15769908

2) Jia Y, Takimoto K. (2006) Mitogen-activated protein kinases control cardiac KChIP2 gene expression. Circ Res. 98(3):386-93. PMID: 16385079

3) Godsel LM, Hsieh SN, Amargo EV, Bass AE, Pascoe-McGillicuddy LT, Huen AC, Thorne ME, Gaudry CA, Park JK, Myung K, Goldman RD, Chew TL, Green KJ. (2005) Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. J Cell Biol. 171(6):1045-59. PMID: 16365169

4) Sirover MA1. (1999) New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta. 1432(2): 159-84. PMID: 10407139

5) Tristan C, Shahani N, Sedlak TW, Sawa A. (2011) The diverse functions of GAPDH: views from different subcellular compartments. Cell Signal. 23(2):317-23. PMID: 20727968

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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NB300-221
Species: Hu, Mu, Rt, Po, Av, Bv, Ca, Ch, ChHa, Dr, Eq, Fe, Ha, Ma-Op, Pm, Rb, Sh, Ze
Applications: WB, Simple Western, ICC/IF, IHC, IP, ChIP, KD, Single-Cell Western

Publications for GAPDH Antibody (NB300-221)(411)

We have publications tested in 16 confirmed species: Human, Mouse, Rat, Bovine, Canine, Chicken, Chinese Hamster, Drosophila, Feline, Opossum, Porcine, Primate, Rabbit, Sheep, Xenopus, Zebrafish.

We have publications tested in 13 applications: B/N, ChIP, Chemotaxis, ICC/IF, IF/IHC, IHC, IHC-Fr, IHC-P, IP, PCR, Simple Western, Single Cell Western, WB.


Filter By Application
B/N
(4)
ChIP
(1)
Chemotaxis
(2)
ICC/IF
(10)
IF/IHC
(2)
IHC
(1)
IHC-Fr
(1)
IHC-P
(1)
IP
(5)
PCR
(1)
Simple Western
(1)
Single Cell Western
(1)
WB
(332)
All Applications
Filter By Species
Human
(161)
Mouse
(113)
Rat
(40)
Bovine
(6)
Canine
(1)
Chicken
(4)
Chinese Hamster
(1)
Drosophila
(1)
Feline
(1)
Opossum
(1)
Porcine
(5)
Primate
(1)
Rabbit
(1)
Sheep
(4)
Xenopus
(1)
Zebrafish
(1)
All Species
Showing Publications 1 - 10 of 411. Show All 411 Publications.
Publications using NB300-221 Applications Species
Steppan J, Wang H, Nandakumar K et al. LOXL2 inhibition ameliorates pulmonary artery remodeling in pulmonary hypertension bioRxiv 2023-10-29 [PMID: 37961202] (WB)

Details:
Dilution 1:5000
WB
Chua B, Ferry L, Domrane C et al. E4F1 and ZNF148 are transcriptional activators of the A57C and wildtypeTERTpromoter bioRxiv 2023-01-15 [PMID: 37918959] (WB, Human) WB Human
Arumugam TV, Alli-Shaik A, Liehn EA et al. Multiomics analyses reveal dynamic bioenergetic pathways and functional remodeling of the heart during intermittent fasting Elife 2023-09-28 [PMID: 37769126]
Kalinin S, Feinstein DL Astrocyte lipocalin-2 modestly effects disease severity in a mouse model of multiple sclerosis while reducing mature oligodendrocyte protein and mRNA expression Neuroscience letters 2023-09-23 [PMID: 37748675] (IHC-P, Mouse)

Details:
1:300 IHC-P dilution
IHC-P Mouse
Guo D, Yu M, Guo H et al. Panax notoginseng saponins inhibits oxidative stress- induced human nucleus pulposus cell apoptosis and delays disc degeneration in vivo and in vitro Journal of ethnopharmacology 2023-09-15 [PMID: 37716491] (WB, Human) WB Human
Yasuhara J, Manivannan SN, Majumdar U et al. Novel pathogenic GATA6 variant associated with congenital heart disease, diabetes mellitus and necrotizing enterocolitis Pediatric research 2023-09-12 [PMID: 37700164]
Huang CL, Chen DY, Tzang CC et al. Celastrol attenuates human parvovirus B19 NS1?induced NLRP3 inflammasome activation in macrophages Molecular Medicine Reports 2023-08-31 [PMID: 37654202] (WB) WB
Cheng W, Lawson C, Liu H et al. Exploration of Mediators Associated with Myocardial Remodelling in Feline Hypertrophic Cardiomyopathy Animals 2023-06-26 [PMID: 37443910] (WB, Feline) WB Feline
Kong MJ, Han SJ, Seu SY et al. Shortening of primary cilia length is associated with urine concentration in the kidneys Kidney Research and Clinical Practice 2023-05-31 [PMID: 37313611] (WB) WB
Natsagdorj A, Izumi K, Hiratsuka K et al. The CCL2-CCR2 Axis Contributes to Migration of Cabazitaxel-resistant Prostate Cancer Cells Anticancer research 2023-06-01 [PMID: 37247914]
Show All 411 Publications.

Reviews for GAPDH Antibody (NB300-221) (19) 4.419

Average Rating: 4.4
(Based on 19 reviews)
We have 19 reviews tested in 6 species: Human, Mouse, Rat, Human and Mouse, Other, cat.

Reviews using NB300-221:
Filter by Applications
WB
(19)
All Applications
Filter by Species
Human
(8)
Mouse
(2)
Rat
(3)
Human and Mouse
(1)
Other
(4)
cat
(1)
All Species
Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Human 06/14/2021
View

Summary

ApplicationWestern Blot
Sample Testedeye tissue
SpeciesHuman
Lot082219

Comments

CommentsUse at 1:500
  5
reviewed by:
Verified Customer
WB Human 01/31/2019
View

Summary

ApplicationWestern Blot
Sample TestedHuman nucleus pulposus cell culture
SpeciesHuman
Lot111713
Western Blot GAPDH NB300-221
Enlarge
5
reviewed by:
Wan-Ching Cheng
WB cat 09/13/2018
View

Summary

ApplicationWestern Blot
Sample TestedHeart tissue
Speciescat
Lot083017
Western Blot GAPDH NB300-221
Enlarge
4
reviewed by:
Verified Customer
WB Human and Mouse 06/29/2017
View

Summary

ApplicationWestern Blot
Sample Tested293FT cell lysate,nih3t3 cell lysate
SpeciesHuman and Mouse
Lot120116
Western Blot GAPDH NB300-221
Enlarge
5
reviewed by:
James Xiao
WB Mouse 05/22/2017
View

Summary

ApplicationWestern Blot
Sample TestedMouse lung endothelial cells
SpeciesMouse
Lot012517

Comments

CommentsCell lysates of treated mouse lung endothelial cells were analyzed by western blotting.
Western Blot GAPDH NB300-221
Enlarge
3
reviewed by:
Verified Customer
WB Human 01/14/2015
View

Summary

ApplicationWestern Blot
Sample TestedHuman nucleus pulposus cell culture
SpeciesHuman
Lot111711
Western Blot GAPDH NB300-221
Enlarge
5
reviewed by:
Verified Customer
WB Other 01/09/2015
View

Summary

ApplicationWestern Blot
Sample TestedChicken Pectoralis Major
SpeciesOther
Lot111711
Western Blot GAPDH NB300-221
Enlarge
5
reviewed by:
Eric England
WB Other 10/08/2014
View

Summary

ApplicationWestern Blot
Sample TestedWhole Skeletal Muscle Lysate
SpeciesOther
Lot111711
Western Blot GAPDH NB300-221
Enlarge
4
reviewed by:
Liyong Zhang
WB Mouse 07/01/2014
View

Summary

ApplicationWestern Blot
SpeciesMouse
Western Blot GAPDH NB300-221
Enlarge
4
reviewed by:
Verified Customer
WB Other 12/16/2013
View

Summary

ApplicationWestern Blot
Sample TestedZebrafish embryo lysates
SpeciesOther
  4
reviewed by:
Verified Customer
WB Human 12/23/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  5
reviewed by:
Verified Customer
WB Human 12/21/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman Hepatoblastoma
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Rat 12/06/2011
View

Summary

ApplicationWestern Blot
Sample TestedRat
SpeciesRat
  4
reviewed by:
Verified Customer
WB Human 12/05/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Rat 11/23/2011
View

Summary

ApplicationWestern Blot
Sample TestedRat brain
SpeciesRat
  4
reviewed by:
Verified Customer
WB Rat 11/21/2011
View

Summary

ApplicationWestern Blot
Sample TestedRat
SpeciesRat
  5
reviewed by:
Verified Customer
WB Human 09/30/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Other 03/25/2010
View

Summary

ApplicationWestern Blot
Sample TestedCHO cells, Sample Amount: 20ug
SpeciesOther
Lot113003
  5
reviewed by:
Verified Customer
WB Human 06/04/2009
View

Summary

ApplicationWestern Blot
Sample TestedEAhy926 cell lysate, Sample Amount: 15ug
SpeciesHuman
LotP1

Product General Protocols

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 GAPDH Antibody (NB300-221). (Showing 1 - 7 of 7 FAQs).

  1. We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
    • GAPDH is expressed in HeLa cell lines but whether or not it is the best option for a loading control antibody depends on the size of the protein you are interested in. GAPDH antibody is a good choice for low to mid MW targets, as GAPDH detects around 35 kDa.
  2. I want to know why GAPDH is used as a loading control antibody in WB techniques.
    • Our GAPDH antibody makes a good loading control because GAPDH is  a housekeeping gene essential to metabolism that is globally expressed in most tissue types and cell lines.
  3. I need a suitable loading control antibody that I can use in western blot of rat nerve extracts, which do you recommend?
    • This GAPDH antibody (NB300-221) is a good choice for a loading control antibody and has been cited in over 180 publications and used on rat lysates with good results.
  4. I'm looking for a loading control between 24kDa - 65kDa for use in simple western, we tried an actin antibody  we already had but it didn't work. What can you recommend?
    • Our GAPDH antibody NB300-221 has been validated in simple western and detects around 35 kDa so I think it would be a good choice for a simple western loading control antibody based on your size requirements.
  5. What secondary antibody should I use for visualization of the GAPDH antibody?
    • This GAPDH antibody is raised in mouse so you would want to use an anti-mouse secondary with the dye of your choice.
  6. Do you offer a smaller size for the GAPDH antibodies?
    • We offer sample sizes for many of our GAPDH loading control antibodies including NB300-221.
  7. I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.
    • For homogenized tissue, beta-actin and GAPDH are both fine.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Additional GAPDH Products

Research Areas for GAPDH Antibody (NB300-221)

Find related products by research area.

Blogs on GAPDH. Showing 1-10 of 13 blog posts - Show all blog posts.

Considerations for Quantitative Western blotting
By Jamshed Arslan, Pharm. D., PhD. Since its inception in 1979, Western blotting has undergone several developments. The use of radioactive probes was common throughout 1980s, but utilizing secondary antibodies labell...  Read full blog post.

The use of Beta Actin (AC-15) as a loading control across multiple species
Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two...  Read full blog post.

Tips on choosing an ideal loading control antibody for Western Blotting
Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. When p...  Read full blog post.

GAPDH - A "Housekeeping" Gene With Diverse Functions in Cellular Homeostasis
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a well-known housekeeping gene with functions in glycolysis. Many biologists are familiar with the gene and use GAPDH antibodies for a loading control when performing western blots. However, this ...  Read full blog post.

A New Standard in Antibody Testing - Simple Western Certified Antibodies
The Western blot is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. The tradi...  Read full blog post.

GAPDH: More than a housekeeping gene
GAPDH is a 146kD tetramer glycolytic pathway metabolic enzyme composed of four 30-40 kDa subunits. It is responsible for reversibly phosphorylating its substrate glyceraldehyde 3-phosphate within the glycolytic pathway.  Apart from its role in gl...  Read full blog post.

GAPDH (Glyceraldehyde 3-Phosphate Dehydrogenase)
GAPDH is a 146 kD tetramer glycolytic pathway metabolic enzyme responsible for reversibly phosphorylating glyceraldehyde 3-phosphate. It may have other possible functions in transcriptional activation. GAPDH is highly expressed due to this housekeepin...  Read full blog post.

Time to Start Actin Like a Reliable 'Housekeeper'!
A growing body of data and studies using actin antibodies supports a view of the actin cytoskeleton of smooth muscle cells as a dynamic structure that plays an integral role in regulating the development of mechanical tension and the material properti...  Read full blog post.

Beta Actin and GAPDH: The Importance of Western Blot Loading Controls
What are Western Blot Loading Controls?Western blotting is an essential technique to probe protein expression in complex cell or tissue lysates. To accurately determine protein expression and interpret Western blot results, it is important to use...  Read full blog post.

The GAPDH Antibody in Western Blot Assays
The loading controls on our antibody database are widely used in gel electrophoresis and Western blotting studies. Products like the GAPDH antibody detect "housekeeping" proteins which are abundantly distributed in cells. This makes them useful for ch...  Read full blog post.

Showing 1-10 of 13 blog posts - Show all blog posts.
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Recent Reviews

4.4
5
9
4
9
3
1
2
0
1
0

Verified Customer
06/14/2021
Application: WB
Species: Human

 
Verified Customer
01/31/2019
Application: WB
Species: Human

Wan-Ching Cheng
09/13/2018
Application: WB
Species: cat

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot