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GAPDH Antibody [Unconjugated]

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Western blot shows lysates of mouse, human, and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat GAPDH/G3PDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) followed ...read more
GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 5 µg/mL for 3 hours at ...read more
GAPDH/G3PDH was detected in immersion fixed paraffin-embedded sections of normal human kidney using Goat Anti-Human/Mouse/Rat GAPDH/G3PDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 8 µg/mL ...read more
GAPDH was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Goat Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 3 µg/mL ...read more
Simple Western lane view shows lysates of human brain (motor cortex) tissue and HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for GAPDH at approximately 43 kDa ...read more
Role of Akt pathway in aliskiren's effects on attenuating collagen expression.A and B demonstrate the results of Western blotting for phospho- and total Akt in hearts of mice treated with various diets. C shows changes ...read more
Effect of prodigiosins on the mRNA and protein expression of HO-1 in gastric tissue.Data (means ± SEM of three assays) were normalized to the levels of GAPDH and expressed as fold induction (log2 scale), relative to ...read more
The DU145FP cells express higher levels of cancer stemness related genes. (A) Transcriptome analyses of prostate cancer stemness-related markers. The Log2 transformed ratio of FPKM values (e.g., FP/WT) are indicated by ...read more
Effects of aliskiren on homocysteine-induced matrix expression in cultured cardiac fibroblasts.Fibroblasts were cultured in the presence or absence of D,L, homocysteine 200 µmol/L for 24 hours after pretreatment with ...read more
Role of Akt pathway in aliskiren's effects on attenuating collagen expression.A and B demonstrate the results of Western blotting for phospho- and total Akt in hearts of mice treated with various diets. C shows changes ...read more
Effects of aliskiren on myocardial matrix gene expression.Real time PCR was performed with ventricular tissue obtained from mice after 12-inducing diet and different doses of aliskiren. Figure 3A demonstrates changes in ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, Simple Western, IHC, ICC/IF
Clonality
Polyclonal
Host
Goat
Conjugate
Unconjugated
Concentration
LYOPH

Order Details

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Catalog# & Formulation Size Price

GAPDH Antibody [Unconjugated] Summary

Immunogen
E. coli-derived recombinant human GAPDH/G3PDH
Met1-Ala150
Accession # P04406
Specificity
Detects human, mouse, and rat GAPDH/G3PDH in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human GAPDH-2 is observed. 
Source
N/A
Isotype
IgG
Clonality
Polyclonal
Host
Goat
Gene
GAPDH
Purity Statement
Antigen Affinity-purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Immunocytochemistry 5-25 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL
Reviewed Applications
Read 4 Reviews rated 5
using
AF5718 in the following applications:

Publications
Read Publications using
AF5718 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
Concentration
LYOPH
Reconstitution Instructions
Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for GAPDH Antibody [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

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) 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 and contains two NAD binding sites (Asp35 and Asn316) with a catalytic region between aa 151‑155. GAPDH contains more than 19 posttranslational modifications, including methylation, deamidation and phosphorylation. One splice variant shows a 10 aa substitution for aa 319‑335. Over amino acid 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 (AF5718)(53)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Hamster.

We have publications tested in 3 applications: Flow Cytometry, Simple Western, Western Blot.


Filter By Application
Flow Cytometry
(1)
Simple Western
(1)
Western Blot
(23)
All Applications
Filter By Species
Human
(13)
Mouse
(8)
Rat
(4)
Hamster
(1)
All Species
Showing Publications 1 - 10 of 53. Show All 53 Publications.
Publications using AF5718 Applications Species
Inwoo Seo, Sung‐Won Kim, Jiyu Hyun, Yu‐Jin Kim, Hyun Su Park, Jeong‐Kee Yoon et al. Enhancing viability and angiogenic efficacy of mesenchymal stem cells via HSP90 alpha and HSP27 regulation based on ROS stimulation for wound healing Bioengineering & Translational Medicine 6/7/2023 [PMID: 37693062]
Lal S, Bhola N H et al. Discovery and Characterization of ZL-2201, a Potent, Highly Selective, and Orally Bioavailable Small-molecule DNA-PK Inhibitor. Cancer Res Commun 2024-09-01 [PMID: 37663435] (Simple Western, Human) Simple Western Human
Matsumoto C, Sekine H, Zhang N et al. Role of p53 in Cisplatin-Induced Myotube Atrophy International Journal of Molecular Sciences 5/24/2023 [PMID: 37298128]

Details:
Dilution: 10 ?g/mL
Kira Engeroff, Davide Warm, Stefan Bittner, Oriane Blanquie Different activity patterns control various stages of Reelin synthesis in the developing neocortex Cerebral Cortex 7/24/2023 [PMID: 37288494]
Singh R, Rossini V, Stockdale SR et al. An IBD-associated pathobiont synergises with NSAID to promote colitis which is blocked by NLRP3 inflammasome and Caspase-8 inhibitors Gut Microbes 12/31/2023 [PMID: 36656595]
Longxiang Huang, Qin Ye, Chunlin Lan, Xiaohui Wang, Yihua Zhu AZD6738 Inhibits fibrotic response of conjunctival fibroblasts by regulating checkpoint kinase 1/P53 and PI3K/AKT pathways Frontiers in Pharmacology 9/20/2022 [PMID: 36204234]
CJ Chen, CC Wu, CY Chang, JR Li, YC Ou, WY Chen, SL Liao, JD Wang Metformin Mitigated Obesity-Driven Cancer Aggressiveness in Tumor-Bearing Mice International Journal of Molecular Sciences, 2022-08-15;23(16):. 2022-08-15 [PMID: 36012397] (Western Blot, Mouse) Western Blot Mouse
Sang Ho Lee, Yu-Jin Kim, Yeong Hwan Kim, Han Young Kim, Suk Ho Bhang Enhancing therapeutic efficacy of human adipose-derived stem cells by modulating photoreceptor expression for advanced wound healing Stem Cell Research & Therapy 12/1/2022 [PMID: 35619187]
W Xiao, A Pinilla-Ba, J Faulkner, X Song, P Prabhakar, H Qiu, KW Moremen, A Ludwig, PJ Dempsey, P Azadi, L Wang Robo4 is constitutively shed by ADAMs from endothelial cells and the shed Robo4 functions to inhibit Slit3-induced angiogenesis Scientific Reports, 2022-03-14;12(1):4352. 2022-03-14 [PMID: 35288626] (Western Blot, Human, Mouse) Western Blot Human, Mouse
Maria Prendecki, Stephen P McAdoo, Tabitha Turner‐Stokes, Ana Garcia‐Diaz, Isabel Orriss, Kevin J Woollard et al. Glomerulonephritis and autoimmune vasculitis are independent of P2RX7 but may depend on alternative inflammasome pathways The Journal of Pathology 7/1/2022 [PMID: 35239186]
Show All 53 Publications.

Reviews for GAPDH Antibody (AF5718) (4) 54

Average Rating: 5
(Based on 4 reviews)
We have 4 reviews tested in 2 species: Mouse, Human and Pig.

Reviews using AF5718:
Filter by Applications
WB
(3)
Simple Western
(1)
All Applications
Filter by Species
Mouse
(3)
Human and Pig
(1)
All Species
Images Ratings Applications Species Date Details
  5
reviewed by:
Verified Customer
WB Mouse 02/23/2022
View

Summary

ApplicationWestern Blot
Sample TestedColon tissue
SpeciesMouse
LotCCSE0421031
Simple Western GAPDH AF5718
Enlarge
5
reviewed by:
Verified Customer
Simple Western Human and Pig 08/05/2019
View

Summary

ApplicationSimple Western
Sample Testedhuman liver cell,Liver tissue,Muscle tissue,Skin tissue
SpeciesHuman and Pig
LotCCSE0418051
  5
reviewed by:
Verified Customer
WB Mouse 01/08/2015
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 22868230
SpeciesMouse
  5
reviewed by:
Verified Customer
WB Mouse 01/08/2015
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 23841780
SpeciesMouse

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 (AF5718). (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

Additional GAPDH Products

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
4
4
0
3
0
2
0
1
0

Verified Customer
02/23/2022
Application: WB
Species: Mouse

Verified Customer
08/05/2019
Application: Simple Western
Species: Human and Pig

 
Verified Customer
01/08/2015
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol GAPDH
Uniprot