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GAPDH Antibody

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Western Blot: GAPDH Antibody [NB300-327] - Representative immunoblot of KDR and FGFR CAM protein expression levels examined by Western blotting. GNS, graphene nanosheet; NG, graphite nanoparticle; ND, diamond ...read more
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody [NB300-327] - Analysis of HeLa cells stained with rabbit pAb to GAPDH, dilution 1:2000 in red. Blue is Hoescht staining of nuclear DNA. The GAPDH antibody produces ...read more
Western Blot: GAPDH Antibody [NB300-327] - Theoretical molecular weight: 36 kDa. Detection of GAPDH in mouse liver.
Western Blot: GAPDH Antibody [NB300-327] - Western blot analysis of extracts from Jurkat and HeLa cells using NB300-327 at 1:1000. Theoretical molecular weight: 36 kDa.
Western Blot: GAPDH Antibody [NB300-327] - Analysis of different cell cytosolic or nuclear enriched fractions, GAPDH antibody, dilution 1:20,000 (Red): [1] protein standard, [2] NIH-3T3 cytosolic, [3] NIH-3T3 nuclear, ...read more
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody [NB300-327] - The GAPDH antibody was tested in Hela cells at a 1:500 dilution against Dylight 488 (Green). Alpha-tubulin and nuclei were counterstained with ...read more
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody [NB300-327] - Confocal immunofluorescence analysis of HeLa cells using GAPDH (NB300-327) antibody (green). Nuclei was counterstained with DAPI (blue).
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody [NB300-327] - IF Confocal analysis of C2C12 cells using GAPDH antibody (NB300-327, 1:20). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary ...read more
Simple Western: GAPDH Antibody [NB300-327] - Simple Western lane view shows a specific band for GAPDH in 0.2 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation ...read more
Western Blot: GAPDH Antibody [NB300-327] - Effects of subchronic administration (7 and 14 days) of therapeutic relevant concentration of Brex (Brex: 300 nM) and interaction between Brex and 5-HT1A receptor (5-HT1AR) ...read more
Representative immunoblot of KDR and FGFR CAM protein expression levels examined by Western blotting. GNS, graphene nanosheet; NG, graphite nanoparticle; ND, diamond nanoparticle; C60, fullerene C60; MWNT, multi-wall ...read more
Growth of lactobacilli on IECs and their influence towards C. albicans cytotoxicity, adhesion, hyphal length and translocation. (G) E-Cadherin protein expression analyzed by western blot compared to GAPDH in IECs that ...read more
HMGB1 regulated autophagy in thyroid cancer cells (a) FTC-133/TPC-1 cells were transfected with HMGB1 shRNA and control shRNA and then starved by HBSS for 2 h. And LC3-I/II level was assayed by Western blot; (b) ...read more
Compound MI-743 causes intracellular 8-oxo-dG accumulation and DNA damage.a MGC-803, HGC-27 and GES-1 cells were treated with DMSO or 5 µM MI-743 for 48 h. Intracellular 8-oxo-dG was stained with Cy3-conjugated ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Ba, Bv, Ch, Eq, Fu, In, YeSpecies Glossary
Applications WB, Simple Western, ICC/IF, IHC
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated

Order Details

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GAPDH Antibody Summary

Immunogen
This GAPDH antibody was developed against full length recombinant human GAPDH
Marker
Cytosolic Marker
Clonality
Polyclonal
Host
Rabbit
Gene
GAPDH
Purity
Unpurified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Immunocytochemistry/ Immunofluorescence 1:500-1:1000
  • Immunohistochemistry 1:10000
  • Simple Western 1:500
  • Western Blot 1:5000
Application Notes
This GAPDH antibody is useful Immunocytochemistry/Immunofluorescence and Western blot. In Western blot a band is observed at approx. 36kDa, and on cells in tissue culture the antibody stains in a punctate cytoplasmic fashion.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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 3 Reviews rated 5
using
NB300-327 in the following applications:

Publications
Read Publications using
NB300-327 in the following applications:

Reactivity Notes

Bacteria reactivity reported in scientific literature (PMID: 31413153). Fungi reactivity reported in scientific literature (PMID:31413153).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Supplied as serum
Preservative
0.035% Sodium Azide
Purity
Unpurified

Alternate Names for GAPDH Antibody

  • aging-associated gene 9 protein
  • EC 1.2.1
  • EC 1.2.1.12
  • EC 2.6.99.-
  • G3PD
  • G3PDH
  • GAPD
  • 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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NB300-327
Species: Hu, Mu, Rt, Po, Ba, Bv, Ch, Eq, Fu, In, Ye
Applications: WB, Simple Western, ICC/IF, IHC

Publications for GAPDH Antibody (NB300-327)(48)

We have publications tested in 8 confirmed species: Human, Mouse, Rat, Bacteria, Chicken, Fungi, Invertebrate, Yeast.

We have publications tested in 4 applications: ICC/IF, In vitro, Simple Western, WB.


Filter By Application
ICC/IF
(1)
In vitro
(1)
Simple Western
(2)
WB
(34)
All Applications
Filter By Species
Human
(16)
Mouse
(7)
Rat
(6)
Bacteria
(1)
Chicken
(3)
Fungi
(1)
Invertebrate
(1)
Yeast
(2)
All Species
Showing Publications 1 - 10 of 48. Show All 48 Publications.
Publications using NB300-327 Applications Species
Loujain Aloui, Elizabeth S. Greene, Travis Tabler, Kentu Lassiter, Kevin Thompson, Walter G. Bottje, Sara Orlowski, Sami Dridi Effect of heat stress on the hypothalamic expression profile of water homeostasis‐associated genes in low‐ and high‐water efficient chicken lines Physiological Reports 2024-03-11 [PMID: 38467563]
Damian M Janecki, Raneet Sen, Natalia Szóstak, Arkadiusz Kajdasz, Martyna Kordyś, Kinga Plawgo, Dmytro Pandakov, Anna Philips, Zbigniew Warkocki LINE-1 mRNA 3′ end dynamics shape its biology and retrotransposition potential Nucleic Acids Research 2024-04-12 [PMID: 38197223]
Drazkowska K, Tomecki R, Warminski M et al. 2'-O-Methylation of the second transcribed nucleotide within the mRNA 5' cap impacts the protein production level in a cell-specific manner and contributes to RNA immune evasion Nucleic Acids Research 2022-09-09 [PMID: 36018811] (WB) WB
Okada M, Fukuyama K, Motomura E Dose-Dependent Biphasic Action of Quetiapine on AMPK Signalling via 5-HT7 Receptor: Exploring Pathophysiology of Clinical and Adverse Effects of Quetiapine International journal of molecular sciences 2022-08-14 [PMID: 36012369] (Simple Western, Rat)

Details:
Dilution used 1:300
Simple Western Rat
Skvorak, KJ. Investigation of gene and cellular therapies to cure maple syrup urine disease (MSUD) in a genetically engineered mouse model. PhD Thesis, University of Pittsburgh. 2008-01-01 (WB, Mouse) WB Mouse
Franklin AL Translation of Partially-Decayed Messenger RNAs in Yeast Thesis (WB, Yeast) WB Yeast
H M Stone, E Unal, T A Romano, P E Turner Beluga whale and bottlenose dolphin ACE2 proteins allow cell entry mediated by spike protein from three variants of SARS-CoV-2. Biology letters 2023-12-07 [PMID: 38053365]
Yang W, Han B, Chen Y, Geng F SAAL1, a novel oncogene, is associated with prognosis and immunotherapy in multiple types of cancer Aging 2022-08-13 [PMID: 35963646] (WB, Human) WB Human
Fukuyama K, Motomura E, Okada M Brexpiprazole Reduces 5-HT7 Receptor Function on Astroglial Transmission Systems International journal of molecular sciences 2022-06-12 [PMID: 35743014] (Simple Western, WB, Rat) Simple Western, WB Rat
Baron DM, Fenton AR, Saez-Atienzar S et al. ALS-associated KIF5A mutations abolish autoinhibition resulting in a toxic gain of function Cell reports 2022-04-05 [PMID: 35385738] (In vitro, Mouse) In vitro Mouse
Show All 48 Publications.

Reviews for GAPDH Antibody (NB300-327) (3) 53

Average Rating: 5
(Based on 3 reviews)
We have 3 reviews tested in 2 species: Human, Mouse.

Reviews using NB300-327:
Filter by Applications
WB
(1)
ICC
(2)
All Applications
Filter by Species
Human
(2)
Mouse
(1)
All Species
Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Human 09/25/2014
View

Summary

ApplicationWestern Blot
Sample TestedCell lysate from HeLa cells
SpeciesHuman
Immunocytochemistry GAPDH NB300-327
Enlarge
5
reviewed by:
Bryan Tinsley
ICC Mouse 06/13/2014
View

Summary

ApplicationImmunocytochemistry
SpeciesMouse
Immunocytochemistry GAPDH NB300-327
Enlarge
5
reviewed by:
Bryan Tinsley
ICC Human 05/21/2014
View

Summary

ApplicationImmunocytochemistry
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 (NB300-327). (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 (NB300-327)

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

5
5
3
4
0
3
0
2
0
1
0

 
Verified Customer
09/25/2014
Application: WB
Species: Human

Bryan Tinsley
06/13/2014
Application: ICC
Species: Mouse

Bryan Tinsley
05/21/2014
Application: ICC
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot