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GAPDH Antibody (6C5cc)

Images

 
Western Blot: GAPDH Antibody (6C5) [NB600-502] - MUS81 promotes DNA synthesis during mitosis in BRCA2-deficient cells. Cell extracts were immunoblotted as indicated. SMC1 and GAPDH were used as loading controls. Image ...read more
Western Blot: GAPDH Antibody (6C5) [NB600-502] - Western blot analysis of GAPDH expression in 1) HeLa, 2) NTERA-2, 3) A-431, 4) HepG2, 5) MCF-7, 6) NIH 3T3, 7) PC-12 and 8) COS-7 whole cell lysates. Theoretical ...read more
Western Blot: GAPDH Antibody (6C5) [NB600-502] - Western blot of FGF23 in the kidney. GAPDH antibody (6C5) served as an internal control. Each value represents the mean +/- SEM. Citation: Sugiura H, Matsushita A, Futaya ...read more
Simple Western: GAPDH Antibody (6C5) [NB600-502] - Simple Western lane view shows a specific band for GAPDH in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa ...read more
Simple Western: GAPDH Antibody (6C5) [NB600-502] - Anaylsis in rat brain and spinal cord. This image includes the GAPDH at the predicted kDa as well as a glucocorticoid receptor antibody (peak 102). Verified customer ...read more
Representative picture of western blot in histopathologically unchanged tissue (N) and primary cancerous tissue (C) from patients with CRC. Immunodetection of bands was performed with Rp anti- PHD1, - PHD2, - PHD3 and - ...read more
Immunostaining of HN30 and MCF-12a cells for TRAF2 3 days post-transfectionMCF-12a NT (DAPI): non-treated MCF-12a DAPI stained, MCF-12a NT (TRAF2-FITC): non-treated MCF-12a FITC stained for TRAF2, MCF-12a pNSP 5a3a ...read more
Baclofen reversed the changes of protein markers characteristic for autophagy in hippocampal CA1 area under chronic cerebral hypoperfusion.(a–c) Five weeks after induction of hypoperfusion, p-mTOR was significantly ...read more
The production of DSBs depends on Top1 degradation in CPT-treated quiescent cells. (A–C) Serum-starved WI38 hTERT cells were co-transfected with siRNAs against cullin 3 and cullin 4B or against a control sequence and ...read more
GFAP expression and synthesis in the ON and ONH in a murine glaucoma model. (A) GFAP immunoreactivity in the glial lamina of 1-month (green, upper panel) and 2 months (green, middle and lower panel) old beta B1-CTGF1 ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Am, Ca, ChHa, Fe, Fi, Rb, Bv(-), Gt(-)Species Glossary
Applications WB, Simple Western, ELISA, ICC/IF, IHC, IP
Clone
6C5cc
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

Order Details

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GAPDH Antibody (6C5cc) Summary

Immunogen
Hybridoma clone has been derived from hybridization of Sp2/0 myeloma cells with spleen cells of Balb/c mice immunized with Rabbit GAPDH.
Localization
Cytoplasmic, Nucleus, Membrane.
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
GAPDH
Purity
Protein A purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • ELISA
  • Immunoassay
  • Immunocytochemistry/ Immunofluorescence 1:10-1:2000
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Simple Western 1:100
  • Western Blot 0.5-1 ug/ml
Application Notes
Use in IHC-P reported in scientific literature (PMID:34496231). Since GAPDH is expressed in all cells it is for example becoming the marker of choice for a loading control in Western Blotting.
See Simple Western Antibody Database for Simple Western validation: tested in HeLa lysate (0.5 mg/ml); antibody dilution of 1:100; separated by size; detects a band at 42 kDa
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
A-431 Whole Cell Lysate
HeLa Whole Cell Lysate
MCF-7 Whole Cell Lysate
GAPDH Overexpression Lysate
HepG2 Whole Cell Lysate
NTERA-2 Whole Cell Lysate
PC-12 Whole Cell Lysate
Reviewed Applications
Read 3 Reviews rated 4.7
using
NB600-502 in the following applications:

Publications
Read Publications using
NB600-502 in the following applications:

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information. Chinese Hamster reactivity reported in scientific literature (PMID: 26115091).

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
PBS (pH 7.4)
Preservative
0.09% Sodium Azide
Purity
Protein A purified

Alternate Names for GAPDH Antibody (6C5cc)

  • aging-associated gene 9 protein
  • anti-GAPDH 6c5
  • EC 1.2.1
  • EC 1.2.1.12
  • EC 2.6.99.-
  • G3PD
  • G3PDH
  • GAPD
  • gapdh 6c5
  • GAPDH
  • glyceraldehyde 3-phosphate dehydrogenase
  • glyceraldehyde-3-phosphate dehydrogenase
  • MGC88685
  • 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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NB600-502
Species: Hu, Mu, Rt, Po, Am, Ca, ChHa, Fe, Fi, Rb, Bv(-), Gt(-)
Applications: WB, Simple Western, ELISA, ICC/IF, IHC, IP

Publications for GAPDH Antibody (NB600-502)(84)

We have publications tested in 6 confirmed species: Human, Mouse, Rat, Amphibian, Chinese Hamster, Rabbit.

We have publications tested in 6 applications: B/N, ICC/IF, IHC-P, IP, Simple Western, WB.


Filter By Application
B/N
(1)
ICC/IF
(2)
IHC-P
(1)
IP
(1)
Simple Western
(1)
WB
(62)
All Applications
Filter By Species
Human
(38)
Mouse
(17)
Rat
(4)
Amphibian
(1)
Chinese Hamster
(1)
Rabbit
(1)
All Species
Showing Publications 1 - 10 of 84. Show All 84 Publications.
Publications using NB600-502 Applications Species
Youn EK, Cho HM, Jung JK et al. Pathologic HDAC1/c-Myc signaling axis is responsible for angiotensinogen transcription and hypertension induced by high-fat diet Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 2023-05-25 [PMID: 37244179] (WB, Mouse) WB Mouse
Torices S, Teglas T, Naranjo O et al. Occludin regulates HIV-1 infection by modulation of the interferon stimulated OAS gene family Research square 2023-01-30 [PMID: 36778388] (WB, Human)

Details:
Dilution used in WB 1:20,000
WB Human
Zhao YM, Sun RS, Duan F et al. Intravitreal slow-release dexamethasone alleviates traumatic proliferative vitreoretinopathy by inhibiting persistent inflammation and M�ller cell gliosis in rabbits International journal of ophthalmology 2023-01-18 [PMID: 36659954] (WB, Rabbit)

Details:
Dilution used in WB 1:1000
WB Rabbit
Osborne OM, Kowalczyk JM, Pierre Louis KD Brain endothelium-derived extracellular vesicles containing amyloid-beta induce mitochondrial alterations in neural progenitor cells Extracellular Vesicles and Circulating Nucleic Acids 2022-12-01 [PMID: 36649440] (WB, Human) WB Human
Eymard ED Role of ETS-1 and histone methylation patterns in rapid recall ability of memory T cells Thesis 2021-01-01
Ridgway SW Diffuse Brain Injury Incites Sexual Differences and Hypothalamic-Pituitary-Adrenal Axis Disruptions Thesis
Lopez KA Therapeutic targeting of histone H3 lysine 36 trimethylation-deficient cancers Thesis 2019-01-01 (WB, Human) WB Human
Miktosz A, tukaszuk B, Baranowski M et al. Co-Stimulation-Induced AP-1 Activity is Required for Chromatin Opening During T Cell Activation bioRxiv 2019-05-23 (WB, Human) WB Human
Dimitrios Kleidonas, Matthias Kirsch, Geoffroy Andrieux, Dietmar Pfeifer, Melanie Boerries, Andreas Vlachos Microglia modulate TNF alpha -mediated synaptic plasticity. Glia 2023-05-19 [PMID: 37208965]
Cao R, Chen P, Wang H et al. Intrafusal-fiber LRP4 for muscle spindle formation and maintenance in adult and aged animals Nature communications 2023-02-10 [PMID: 36765071] (WB, Mouse) WB Mouse
Show All 84 Publications.

Reviews for GAPDH Antibody (NB600-502) (3) 4.73

Average Rating: 4.7
(Based on 3 reviews)
We have 3 reviews tested in 2 species: Human, Rat.

Reviews using NB600-502:
Filter by Applications
WB
(2)
Simple Western
(1)
All Applications
Filter by Species
Human
(2)
Rat
(1)
All Species
Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Human 02/05/2022
View

Summary

ApplicationWestern Blot
Sample TestedEpidermoid carcinoma cell line A-431
SpeciesHuman
Lot6C5cc
Simple Western GAPDH NB600-502
Enlarge
4
reviewed by:
Caitlin Hair
Simple Western Rat 06/04/2018
View

Summary

ApplicationSimple Western
Sample Testedbrain and spinal cord
SpeciesRat
Lot16/06-G4-C5

Comments

CommentsWe really like how reliable this antibody is however we have noticed that the baseline on our Wes plates has been slightly elevated.
Western Blot GAPDH NB600-502
Enlarge
5
reviewed by:
Verified Customer
WB Human 06/17/2014
View

Summary

ApplicationWestern Blot
Sample Tested293T and HepG2 lysates
SpeciesHuman

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 (NB600-502). (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 (NB600-502)

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

4.7
5
2
4
1
3
0
2
0
1
0

Verified Customer
02/05/2022
Application: WB
Species: Human

Caitlin Hair
06/04/2018
Application: Simple Western
Species: Rat

Verified Customer
06/17/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot