GAPDH Antibody (686613) [Unconjugated]

Images

 
Western blot shows lysates of human brain tissue, mouse brain tissue, and rat brain tissue. PVDF Membrane was probed with 0.05 µg/mL of Mouse Anti-Human/Mouse/Rat GAPDH/G3PDH Monoclonal Antibody (Catalog # MAB5718) ...read more
GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat GAPDH Monoclonal Antibody (Catalog # MAB5718) at 8 µg/mL for 3 hours at room temperature. ...read more
GAPDH/G3PDH was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-Human/Mouse/Rat GAPDH/G3PDH Monoclonal Antibody (Catalog # MAB5718) at 15 µg/mL overnight at 4 °C. Before ...read more
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and C2C12 mouse myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for GAPDH/G3PDH at approximately 41 kDa (as ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, Simple Western, IHC, ICC/IF
Clone
686613
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

Order Details

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Catalog# & Formulation Size Price

GAPDH Antibody (686613) [Unconjugated] Summary

Immunogen
E. coli-derived recombinant human GAPDH/G3PDH
Met1-Ala150
Accession # P04406
Specificity
Detects human, mouse and rat GAPDH/G3PDH in Western blots. In direct ELISAs, 100% cross-reactivity with recombinant mouse GAPDH/G3PDH and no cross-reactivity with recombinant human GAPDH-2 is observed.
Source
N/A
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
GAPDH
Purity Statement
Protein A or G purified from hybridoma culture supernatant
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 8-25 ug/mL
  • Simple Western 0.25 - 1 ug/mL
  • Western Blot 0.05 ug/mL
Application Notes
In Simple Western only 10-15 uL of the recommended dilution is used per data point.
Reviewed Applications
Read 1 Review rated 5
using
MAB5718 in the following applications:

Publications
Read Publications using
MAB5718 in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Preservative
No Preservative
Reconstitution Instructions
Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for GAPDH Antibody (686613) [Unconjugated]

  • 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

GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) is a 36-40 kDa member of the GAPDH family of enzymes. It is a widely expressed heterotetramer that is found in both the nucleus and cytoplasm. Although GAPDH was initially identified as a glycolytic enzyme that converted G3P into 1,3 diphosphoglycerate, it is now recognized to participate in no less than endocytosis, membrane fusion, vesicular secretory transport, DNA replication and repair, and apoptosis. Human GAPDH is 335 amino acids (aa) in length. There are two NAD binding sites (Asp35 and Asn316) with a catalytic region between aa 151-155. GAPDH contains no fewer that 19 posttranslational modifications, including methylation, deamidation and phosphorylation. One splice variant shows a 10 aa substitution for aa 319‑335. Over
aa 1‑150, human GAPDH shares 92% aa identity with mouse GAPDH.

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 (MAB5718)(13)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 3 applications: Co-Immunoprecipitation, ICC, Western Blot.


Filter By Application
Co-Immunoprecipitation
(1)
ICC
(2)
Western Blot
(10)
All Applications
Filter By Species
Human
(9)
Mouse
(3)
Rat
(1)
All Species
Showing Publications 1 - 10 of 13. Show All 13 Publications.
Publications using MAB5718 Applications Species
F Schneider, I Metz, S Khudayberd, MB Rust Functional Redundancy of Cyclase-Associated Proteins CAP1 and CAP2 in Differentiating Neurons Cells, 2021;10(6):. 2021 [PMID: 34204261] (Western Blot, Mouse) Western Blot Mouse
LJ Kepser, S Khudayberd, LS Hinojosa, C Macchi, M Ruscica, E Marcello, C Culmsee, R Grosse, MB Rust Cyclase-associated protein 2 (CAP2) controls MRTF-A localization and SRF activity in mouse embryonic fibroblasts Scientific Reports, 2021;11(1):4789. 2021 [PMID: 33637797] (Western Blot, Mouse) Western Blot Mouse
J Fisher, OE Sørensen, AHA Abu-Humaid Using chemiluminescence imaging of cells (CLIC) for relative protein quantification Sci Rep, 2020;10(1):18280. 2020 [PMID: 33106566] (ICC, Human) ICC Human
E Cienfuegos, TR Ibarra-Riv, AL Saucedo, LA Ramírez-Ma, D Esquivel-F, I Domínguez-, KJ Alcántara-, DP Moreno-Peñ, G Alarcon-Ga, DR Rodríguez-, L Torres-Gon, LE Muñoz-Espi, E Pérez-Rodr, P Cordero-Pé Effect of sodium (S)-2-hydroxyglutarate in male, and succinic acid in female Wistar rats against renal ischemia-reperfusion injury, suggesting a role of the HIF-1 pathway PeerJ, 2020;8(0):e9438. 2020 [PMID: 32728491] (Western Blot, Rat) Western Blot Rat
G Kaufman, NM Kiburi, D Skrtic The self-renewal dental pulp stem cell microtissues challenged by a toxic dental monomer Biosci. Rep., 2020;40(6):. 2020 [PMID: 32495822] (ICC, Human) ICC Human
KV Lithgow, B Church, A Gomez, E Tsao, S Houston, LA Swayne, CE Cameron Identification of the Neuroinvasive Pathogen Host Target, LamR, as an Endothelial Receptor for the Treponema pallidum Adhesin Tp0751 mSphere, 2020;5(2):. 2020 [PMID: 32238570] (Co-Immunoprecipitation, Human) Co-Immunoprecipitation Human
S Varshney, HX Wei, F Batista, M Nauman, S Sundaram, K Siminovitc, A Tanwar, P Stanley A modifier in the 129S2/SvPasCrl genome is responsible for the viability of Notch1[12f/12f] mice BMC Dev. Biol., 2019;19(1):19. 2019 [PMID: 31590629] (Western Blot, Mouse) Western Blot Mouse
K Watanabe, M Suzukawa, S Arakawa, K Kobayashi, S Igarashi, H Tashimo, H Nagai, S Tohma, T Nagase, K Ohta Leptin enhances cytokine/chemokine production by normal lung fibroblasts by binding to leptin receptor Allergol Int, 2019;0(0):. 2019 [PMID: 31029506] (Western Blot, Human) Western Blot Human
RM Fliefel, SA Entekhabi, M Ehrenfeld, S Otto Geranylgeraniol (GGOH) as a Mevalonate Pathway Activator in the Rescue of Bone Cells Treated with Zoledronic Acid: An In Vitro Study Stem Cells Int, 2019;2019(0):4351327. 2019 [PMID: 30728841] (Western Blot, Human) Western Blot Human
A Sharma, S Bandyopadh, K Chowdhury, T Sharma, R Maheshwari, A Das, G Chakrabart, V Kumar, CC Mandal Metformin exhibited anticancer activity by lowering cellular cholesterol content in breast cancer cells PLoS ONE, 2019;14(1):e0209435. 2019 [PMID: 30625181] (Western Blot, Human) Western Blot Human
Show All 13 Publications.

Review for GAPDH Antibody (MAB5718) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Mouse.

Reviews using MAB5718:
Filter by Applications
WB
(1)
All Applications
Filter by Species
Mouse
(1)
All Species
Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Mouse 02/25/2015
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 24040198
SpeciesMouse

Blocking

Blocking DetailsSee PMID 24040198

Details

Detection NotesSee PMID 24040198

Comments

CommentsIndependent review from antibodyresource.comPublished in PMID 24040198

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for GAPDH Antibody (MAB5718). (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.

Secondary Antibodies

 

Isotype Controls

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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

5
5
1
4
0
3
0
2
0
1
0

 
Verified Customer
02/25/2015
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol GAPDH
Uniprot