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GAPDH Antibody (1A10) - BSA Free

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Western Blot: GAPDH Antibody (1A10) [NBP1-47339] - Analaysis using anti-GAPDH mAb against Hela (1), A549 (2), A431 (3), MCF-7 (4), K562 (5), Jurkat (6), HL60 (7), SKN-SH (8) and SKBR-3 (9) cell lysate. Theoretical ...read more
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody (1A10) [NBP1-47339] - Analysis of methanol-fixed HepG2 (left) and HeLa (right) cells using anti-GAPDH mAb (green), showing cytoplasmic localization. DRAQ5 ...read more
Flow Cytometry: GAPDH Antibody (1A10) [NBP1-47339] - An intracellular stain was performed on U-87 cells with GAPDH [1A10] Antibody NBP1-47339AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% ...read more
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody (1A10) [NBP1-47339] - GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using 2 ug/mL of mouse anti-GAPDH monoclonal ...read more
Immunohistochemistry-Paraffin: GAPDH Antibody (1A10) [NBP1-47339] - Analysis of paraffin-embedded human breast carcinoma (left) and kidney carcinoma (right), showing cytoplasmic localization using anti-GAPDH mAb with ...read more
Simple Western: GAPDH Antibody (1A10) [NBP1-47339] - Lane view shows a specific band for GAPDH in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation ...read more

Product Details

Summary
Reactivity Hu, Mu(-), Rt(-)Species Glossary
Applications WB, Simple Western, ELISA, Flow, ICC/IF, IHC
Clone
1A10
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Format
BSA Free
Concentration
1.0 mg/ml

Order Details

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GAPDH Antibody (1A10) - BSA Free Summary

Immunogen
GAPDH antibody developed against purified recombinant fragment of human GAPDH expressed in E. coli. [UniProt# P04406]
Localization
Cytoplasmic
Marker
Cytosolic Marker
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
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
  • ELISA 1:10000
  • Flow Cytometry 5 ug/ml
  • Immunocytochemistry/ Immunofluorescence 1:200-1:1000
  • Immunohistochemistry 1:200-1:1000
  • Immunohistochemistry-Paraffin 1:200-1:1000
  • Simple Western 1:50
  • Western Blot 1:500-1:2000
Application Notes
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
Jurkat Whole Cell Lysate
A-431 Whole Cell Lysate
HeLa Whole Cell Lysate
MCF-7 Whole Cell Lysate
Reviewed Applications
Read 2 Reviews rated 4.5
using
NBP1-47339 in the following applications:

Publications
Read Publications using
NBP1-47339 in the following applications:

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 25833141).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.03% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for GAPDH Antibody (1A10) - BSA Free

  • 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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Publications for GAPDH Antibody (NBP1-47339)(11)

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

We have publications tested in 3 applications: ICC/IF, WB, Western Blot.


Filter By Application
ICC/IF
(1)
WB
(5)
Western Blot
(1)
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Filter By Species
Human
(6)
Mouse
(1)
All Species
Showing Publications 1 - 10 of 11. Show All 11 Publications.
Publications using NBP1-47339 Applications Species
Drake RR, Louey S, Thornburg KL Maturation of Lipid Metabolism in the Fetal and Newborn Sheep Heart American journal of physiology. Regulatory, integrative and comparative physiology 2023-10-23 [PMID: 37867472]
Bhaoighill MN, Falcón-Pérez JM, Royo F et al. Tuberous Sclerosis Complex cell-derived EVs have an altered protein cargo capable of regulating their microenvironment and have potential as disease biomarkers Journal of extracellular vesicles 2023-06-01 [PMID: 37337371] (WB, Human) WB Human
Ercan H, Resch U, Hsu F et al. A Practical and Analytical Comparative Study of Gel-Based Top-Down and Gel-Free Bottom-Up Proteomics Including Unbiased Proteoform Detection Cells 2023-02-26 [PMID: 36899884] (Western Blot, Human) Western Blot Human
Belardin LB, LEgarE C, Sullivan R et al. Expression of the pro-inflammatory P2Y14 receptor in the non-vasectomized and vasectomized human epididymis Andrology 2022-08-27 [PMID: 36029226] (WB, Human) WB Human
Prifti DK, Lauzier A, Elowe S A commercial ARHGEF17/TEM4 antibody cross-reacts with Nuclear Mitotic Apparatus protein 1 (NuMA) PloS one 2022-07-01 [PMID: 35776709] (ICC/IF, Human) ICC/IF Human
Vitry J, Par� G, Murru A et al. Regulation of the Expression, Oligomerisation and Signaling of the Inhibitory Receptor CLEC12A by Cysteine Residues in the Stalk Region International Journal of Molecular Sciences 2021-09-22 [PMID: 34638548]
Hoesel B, Mussbacher M, Dikorman B, Salzmann M. Androgen receptor dampens tissue factor expression via NF-kB and EGR1. J. Thromb. Haemost. 2018-02-10 [PMID: 29427323] (Human) Human
Li Y, Zhang DW, Lin DQ, Cao LQ. Peroxisome proliferator-activated receptor-gamma inhibits pancreatic cancer cell invasion and metastasis via regulating MMP-2 expression through PTEN. Mol Med Rep. 2015-08-12 [PMID: 26299428] (WB) WB
Moscovitz O, Ben-Nissan G et al. The Parkinson's-associated protein DJ-1 regulates the 20S proteasome. Nat Commun 2015-04-04 [PMID: 25833141] (WB, Mouse) WB Mouse
Zhang Z, Jiang T, Li Q et al. Nodal activates smad and extracellular signal-regulated kinases 1/2 pathways promoting renal cell carcinoma proliferation. Mol Med Rep 2015-07-01 [PMID: 25672326]
Show All 11 Publications.

Reviews for GAPDH Antibody (NBP1-47339) (2) 4.52

Average Rating: 4.5
(Based on 2 reviews)
We have 2 reviews tested in 1 species: Human.

Reviews using NBP1-47339:
Filter by Applications
WB
(2)
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Filter by Species
Human
(2)
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Images Ratings Applications Species Date Details
  4
reviewed by:
Verified Customer
WB Human 12/23/2014
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Summary

ApplicationWestern Blot
Sample TestedSee PMID 24421388
SpeciesHuman
Western Blot GAPDH NBP1-47339
Enlarge
5
reviewed by:
Ranveer Singh
WB Human 06/02/2014
View

Summary

ApplicationWestern Blot
Sample TestedHeLa cells
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 (NBP1-47339). (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 (NBP1-47339)

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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 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

4.5
5
1
4
1
3
0
2
0
1
0

 
Verified Customer
12/23/2014
Application: WB
Species: Human

Ranveer Singh
06/02/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol GAPDH
Entrez
Uniprot