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Monoclonal Antibody to GAPDH - Loading Control |
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| Catalog No: |
IMG-5019A-1 |
| Contents: |
50 ul antibody solution containing 50% glycerol, 0.01% thimerosal and 1.0 mg/ml BSA. |
| Isotype: |
Mouse-IgG1 |
| Clone: |
GAPDH 1D4 |
| Species: |
Bovine, Chicken, Human, Mouse, Pig, Rabbit, Rat |
| Host: |
Mouse |
| Application: |
IF/ICC: 1:100-1:1000
IP: see Kang et al
Western blot analysis: 1:1000-1:5000 |
| Storage: |
Store this loading control antibody at -20°C. Avoid multiple freeze/thaw cycles. |
Recommended
Positive Control: |
Most cell lines and tissues |
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Background
Glyceraldehyde 3-Phosphate Dehydrogenase (G3PDH or GPDH) is one of the key enzymes involved in glycolysis; it catalyzes the reversible oxidative phosphorylation of glyceraldehydes-3-phosphate. The gene is constitutively and stably expressed at high levels in almost all tissues and cells, and as such is considered to be a “housekeeping” gene. Housekeeping proteins are useful as a loading control for western blots and protein normalization. They are also useful for visualizing cells in microscopy assays. |
Antigen
Purified pig glyceraldehyde 3-phosphate dehydrogenase enzyme. |
Application Notes
The loading control clone 1D4 has been shown to react with human, cow, pig, mouse, rat, and chicken GAPDH. It is highly conserved and likely that the antibody reacts with additional species. |
Genebank Info (Protein)
NP_002037 |
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Application of the glyceraldehyde 3-phosphate dehydrogenase mAb as a protein loading control in Western blots. Total proteins from various human cell lysates were normalized using the loading control. The blot shown is IMGENEX's INSTA-Blot: Human Cell Lines, Cat. no. IMB-105. |
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Application of the glyceraldehyde 3-phosphate dehydrogenase antibody to visualize cells by fluorescence microscopy. Sh-SY5Y human neuroblastoma cells were stained with IMG-5091A, followed by an FITC labeled secondary (green) and counterstained with Hoechst dye (blue). |
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Application of the glyceraldehyde 3-phosphate dehydrogenase mAb as a protein loading control in Western blots. Total proteins from various rat tissue lysates were normalized using the loading control. Rat tissue lysates, Cat nos: Ovary (40151), Stomach (40130), Lung (40126), Liver (40125), and Heart (40122). |
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Reference
1. Robert D. Barber , Dan W. Harmer , Robert A. Coleman and Brian J. Clark (2005). "GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues". Physiological Genomics 21 (3): 389–395.
2. A. Tarze, A. Deniaud, M. Le Bras, E. Maillier, D. Molle, N. Larochette, N. Zamzami, G. Jan, G. Kroemer, and C. Brenner (2007). GAPDH, a novel regulator of the pro-apoptotic mitochondrial membrane permeabilization. Oncogene 26 (18): 2606–2620.
3. Georgina M. Aldridge, David M. Podrebarac, William T. Greenough, and Ivan Jeanne Weiler. The use of total protein stains as loading controls: an alternative to high-abundance single protein controls in semi-quantitative immunoblotting. J Neurosci Methods. 2008 July 30; 172(2): 250–254.
4. Romero-Calvo I, Ocón B, Martínez-Moya P, Suárez MD, Zarzuelo A, Martínez-Augustin O, de Medina FS. Reversible Ponceau staining as a loading control alternative to actin in Western blots. Anal Biochem. 2010 Jun 15;401(2):318-20.
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Research purposes only.
Not for diagnostic or in vivo use. This product is guaranteed to perform as indicated on the datasheet for one year from the date of purchase.
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