GAPDH Antibody

Images

 
Western Blot: GAPDH Antibody [NB100-56875] - Analysis of GAPDH using this antibody at 1:500 in nuclear and cytoplasmic fractions made from Drosophila head extracts. Data courtesy of Dr. Jerry Lin, University of ...read more
Western Blot: GAPDH Antibody [NB100-56875] - Analysis of GAPDH in the multiple human tumor cell line lysate INSTA-Blot using this antibody. 25 ug/ml. Theoretical molecular weight: 36 kDa.
Western Blot: GAPDH Antibody [NB100-56875] - Analysis using HeLa cells. Theoretical molecular weight: 36 kDa.
Western Blot: GAPDH Antibody [NB100-56875] - Vps13 co-fractionates with Rab7 and Rab5. Western blot analysis of control fly head samples fractionated into a cytosolic and membrane fraction from postnuclear supernatant ...read more
Western Blot: GAPDH Antibody [NB100-56875] - Particle-induced Changes in Apoptosis are Restored by Firoin. Peripheral blood neutrophils from healthy donors (n57), 2 h pre-treated with indicated amounts of firoin (F) or ...read more
Simple Western: GAPDH Antibody [NB100-56875] - Lane view shows a specific band for GAPDH in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing A246conditions using the 12-230 kDa separation system. ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Ca, Dr, Fe, Ha, Pm, Po, MaSpecies Glossary
Applications WB, Simple Western, IB, ICC/IF, IHC
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Order Details

GAPDH Antibody Summary

Immunogen
Amino acids 73-87 PITIFQERDPSKIKW of glyceraldehyde 3-phosphate dehydrogenase protein were used as the immunogen of this GAPDH antibody.
Marker
Cytosolic Marker
Predicted Species
Porcine (93%), Gerbil (93%). Backed by our 100% Guarantee.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
GAPDH
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:500-1:2000
  • Simple Western 1:500
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
Application Notes
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size. Use in immunoblotting reported in scientific literature (PMID: 28545464). Use in ICC/IF was reported in scientific literature (PMID: 31312260). Use in Immunohistochemistry reported in scientific literature (PMID:32123074).
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
HeLa Whole Cell Lysate
Human Lung Whole Tissue Lysate (Adult Whole Normal)
Reviewed Applications
Read 5 Reviews rated 4.8
using
NB100-56875 in the following applications:

Publications
Read Publications using
NB100-56875 in the following applications:

  • IB
    2 publications
  • 1 publication
  • IHC
    1 publication
  • WB
    68 publications

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: 100% homologous in baboon, chimp and macaque; salamander(86%).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

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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Supplier Logo

Publications for GAPDH Antibody (NB100-56875)(90)

We have publications tested in 10 confirmed species: Human, Mouse, Rat, Canine, Drosophila, Drosophila , Feline, Hamster, Porcine, Primate.

We have publications tested in 4 applications: IB, ICC/IF, IHC, WB.


Filter By Application
IB
(2)
ICC/IF
(1)
IHC
(1)
WB
(68)
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Human
(41)
Mouse
(15)
Rat
(12)
Canine
(1)
Drosophila
(3)
Drosophila
(1)
Feline
(1)
Hamster
(1)
Porcine
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 90. Show All 90 Publications.
Publications using NB100-56875 Applications Species
Heinonen I, Sorop O, van Dalen BM Cellular, mitochondrial and molecular alterations associate with early left ventricular diastolic dysfunction in a porcine model of diabetic metabolic derangement Sci Rep Aug 6 2020 [PMID: 32764569] (WB, Porcine) WB Porcine
Casselbrant A, FAndriks L, Wallenius V Glycocholic acid and butyrate synergistically increase vitamin D-induced calcium uptake in Caco-2 intestinal epithelial cell monolayers Bone Rep Dec 1 2020 [PMID: 32715032] (WB, Human) WB Human
Xie J, Yuan S, Peng L et al. Antitumor immunity targeting fibroblast activation protein‑ alpha in a mouse Lewis lung carcinoma model Oncol Lett May 18 2020 [PMID: 32566014] (WB, Mouse) WB Mouse
Rosen E, Kryndushkin D, Aryal B et al. Acute total body ionizing gamma radiation induces long-term adverse effects and immediate changes in cardiac protein oxidative carbonylation in the rat PLoS ONE Jun 4 2020 [PMID: 32497067]
Takizawa M, Nakano M, Fukami T, Nakajima M Decrease in ADAR1 expression by exposure to cigarette smoke enhances susceptibility to oxidative stress Toxicol. Lett. May 18 2020 [PMID: 32439581] (Human) Human
Yang JE, Rossignol ED, Chang D et al. Complexity and ultrastructure of infectious extracellular vesicles from cells infected by non-enveloped virus Sci Rep May 14 2020 [PMID: 32409751] (WB, Human) WB Human
Woodling NS, Aleyakpo B, Dyson MC et al. The neuronal receptor tyrosine kinase Alk is a target for longevity Aging Cell Apr 15 2020 [PMID: 32291952] (WB, Drosophila) WB Drosophila
Ondo K,Arakawa H,Nakano M et Al. SLC35B1 significantly contributes to the uptake of UDPGA into the endoplasmic reticulum for glucuronidation catalyzed by UDP-glucuronosyltransferases Biochem. Pharmacol. Mar 14 2020 [PMID: 32179043] (WB, Human) WB Human
Fong Ly, Taccioli C, Palamarchuk A et Al. Abrogation of esophageal carcinoma development in miR-31 knockout rats Proc. Natl. Acad. Sci. U.S.A. Mar 17 2020 [PMID: 32123074] (IHC, Human) IHC Human
Casella A, Long C, Zhou J et Al. Differential Frequency of CYP2R1 Variants Across Populations Reveals Pathway Selection for Vitamin D Homeostasis J. Clin. Endocrinol. Metab. May 1 2020 [PMID: 32115644]
Show All 90 Publications.

Reviews for GAPDH Antibody (NB100-56875) (5) 4.85

Average Rating: 4.8
(Based on 5 reviews)
We have 5 reviews tested in 3 species: Human, Chinese Hamster, Feline.

Reviews using NB100-56875:
Filter by Applications
WB
(5)
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Filter by Species
Human
(2)
Chinese Hamster
(1)
Feline
(1)
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Images Ratings Applications Species Date Details
Western Blot GAPDH NB100-56875
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reviewed by:
Anonymous
WB Human 01/08/2019
View

Summary

ApplicationWestern Blot
Sample Tested3 human pancreas cancer cell lines
SpeciesHuman
Lot04231895B-10
Western Blot GAPDH NB100-56875
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reviewed by:
Anonymous
WB Chinese Hamster 10/13/2017
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Summary

ApplicationWestern Blot
Sample Testedprotein extracts from Lec-2 cells
SpeciesChinese Hamster
Lot04231895B-10

Comments

CommentsI used GAPDH antibody NB100-56875 for Western blot on Lec-2 cells extracts before. It worked really well. Last lot, 04231895B-10, does not recognize hamster GAPDH. I cannot blame Novus since it is polyclonal antibody and they also did not claim that it recognizes hamster GAPDH.
Western Blot GAPDH NB100-56875
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reviewed by:
Anonymous
WB Feline 12/09/2016
View

Summary

ApplicationWestern Blot
Sample TestedHeart lysates
SpeciesFeline
Western Blot GAPDH NB100-56875
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reviewed by:
Anonymous
WB Human 09/01/2015
View

Summary

ApplicationWestern Blot
SpeciesHuman
FileView PDF
 
reviewed by:
Matt Stratton
WB 05/20/2014
View

Summary

ApplicationWestern Blot

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 (NB100-56875). (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.

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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 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 loading controls. A loading control anti...  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

4.8
5
4
4
1
3
0
2
0
1
0

Anonymous
01/08/2019
Application: WB
Species: Human

Anonymous
10/13/2017
Application: WB
Species: Chinese Hamster

Anonymous
12/09/2016
Application: WB
Species: Feline

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot