Mouse GAPDH Monoclonal Antibody | anti-GAPDH antibody

GAPDH (Glyceraldehyde-3-phosphate dehydrogenase, EC 1.2.1.12, GAPD, CDABP0047, OK/SW-cl.12) (Biotin)

Cat. Num. AAA6121096

• Gene Names: GAPDH; G3PD; GAPD; HEL-S-162eP
• Reactivity: Human, Mouse, Rat
• Applications: ELISA, Immunofluorescence, Immunohistochemistry, Western Blot
• Purity: Purified by Protein G Affinity Chromatography
• Synonyms: GAPDH; Monoclonal Antibody; GAPDH (Glyceraldehyde-3-phosphate dehydrogenase; EC 1.2.1.12; GAPD; CDABP0047; OK/SW-cl.12) (Biotin); anti-GAPDH antibody

• Host: Mouse
• Reactivity: Human, Mouse, Rat
• Clonality: Monoclonal
• Isotype: IgG
• Clone Number: 10i46
• Specificity: Recognizes human GAPDH. Species Crossreactivity: mouse, rat
• Purity/Purification: Purified by Protein G Affinity Chromatography
• Form/Format: Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with Biotin.

• Applicable Applications for anti-GAPDH antibody: ELISA (EIA), Immunofluorescence (IF), Immunohistochemistry (IHC) Paraffin, Western Blot (WB)
• Application Notes: Applications are based on unconjugated antibody.
• Immunogen: GAPDH recombinant protein.
• Conjugate: Biotin
• Preparation and Storage: Store product at 4 degree C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20 degree C. Aliquots are stable at -20 degree C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Western Blot (WB)

(Western blot analysis of CEM cell line lysates (35ug/lane). using. GAPDH (arrow) was detected using)

Testing Data

(All lanes : at 1:1000 dilution Lane 1: A431 whole cell lysates Lane 2: C6 whole cell lysates Lane 3: Hela whole cell lysates Lane 4: HUVEC whole cell lysates Lysates/proteins at 20ug/ lane. Secondary IgG (HRP) GtxMoPredicted band size : 36 kD Blocking/Dilution buffer: 5% NFDM/TBST.)

Testing Data

(All lanes : at 1:8000 dilution Lane 1: Hela whole cell lysates Lane 2: A549 whole cell lysates Lane 3: COS-7 whole cell lysates Lane 4: mouse brain lysates Lane 5: C6 whole cell lysates Lane 6: NIH/3T3 whole cell lysates Lysates/proteins at 20ug/ lane. Secondary IgG (HRP) GtxMoPredicted band size : 36 kD Blocking/Dilution buffer: 5% NFDM/TBST.)

Immunohistochemistry (IHC)

(Immunohistochemical analysis of paraffin-embedded H.kidney section using at 1:25 dilution. IgG (HRP) GtxMo was used as the secondary antibody, followed by DAB staining.)

Immunofluorescence (IF)

(Immunofluorescence of Hela cells stained with at 1:25 dilution. An Alexa Fluor® 488-conjugated goat anti-mouse lgG was used as the secondary antibody (green). Cytoplasmic actin was counterstained with Alexa Fluor® 555 conjugated with Phalloidin (red))

Product Categories/Family for anti-GAPDH antibody

Antibodies; Enzymes; Dehydrogenase

References

1. Blatnik,M., Ann. N. Y. Acad. Sci. 1126, 272-275 (2008). 2. Loecken,E.M.Chem. Res. Toxicol. 21 (2), 453-458 (2008). 3. Allen R.W.,J. Biol. Chem. 262:649-653(1987).

NCBI and Uniprot Product Information

• NCBI GI #: 7669492
• NCBI GeneID: 2597
• NCBI Accession #: NP_002037
• NCBI GenBank Nucleotide #: NM_002046.5
• UniProt Accession #: P04406; P00354; Q53X65; E7EUT4
• Molecular Weight: Predicted: 37 kDa; ; Observed: 36 kDa
• NCBI Official Full Name: glyceraldehyde-3-phosphate dehydrogenase isoform 1
• NCBI Official Synonym Full Names: glyceraldehyde-3-phosphate dehydrogenase
• NCBI Official Symbol: GAPDH
• NCBI Official Synonym Symbols: G3PD; GAPD; HEL-S-162eP
• NCBI Protein Information: glyceraldehyde-3-phosphate dehydrogenase; aging-associated gene 9 protein; peptidyl-cysteine S-nitrosylase GAPDH; epididymis secretory sperm binding protein Li 162eP
• UniProt Protein Name: Glyceraldehyde-3-phosphate dehydrogenase
• UniProt Gene Name: GAPDH
• UniProt Synonym Gene Names: GAPD; GAPDH
• UniProt Entry Name: G3P_HUMAN

NCBI Description

This gene encodes a member of the glyceraldehyde-3-phosphate dehydrogenase protein family. The encoded protein has been identified as a moonlighting protein based on its ability to perform mechanistically distinct functions. The product of this gene catalyzes an important energy-yielding step in carbohydrate metabolism, the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). The encoded protein has additionally been identified to have uracil DNA glycosylase activity in the nucleus. Also, this protein contains a peptide that has antimicrobial activity against E. coli, P. aeruginosa, and C. albicans. Studies of a similar protein in mouse have assigned a variety of additional functions including nitrosylation of nuclear proteins, the regulation of mRNA stability, and acting as a transferrin receptor on the cell surface of macrophage. Many pseudogenes similar to this locus are present in the human genome. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Nov 2014]

Uniprot Description

GAPDH: a multifunctional enzyme with both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities. A key glycolytic enzyme that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. An important enzyme for energy metabolism, and the production of ATP and pyruvate through anaerobic glycolysis in the cytoplasm. Additionally, it participates in apoptosis, membrane trafficking, iron metabolism, nuclear activities and receptor mediated cell signaling. Its subcellular localization changes reflecting its multiple activities. Is cytosolic, but is also localized in the membrane, the nucleus, polysomes, the ER and the Golgi. Participates in transcription, RNA transport, DNA replication and apoptosis. S-nitrosylation on Cys-152 following apoptotic stimulates its interaction with SIAH2, which in turn moderates its translocation into the nucleus. Mediates cysteine S-nitrosylation of nuclear target proteins including SIRT1, HDAC2 and DNA-PK. Deregulated in lung cancer, renal cancer, breast cancer, gastric cancer, glioma, liver cancer, colorectal cancer, melanoma, prostatic cancer, pancreatic cancer and bladder cancer. Its increased expression and enzymatic activity is associated with cell proliferation and tumorigenesis, Oxidative stress impairs GAPDH catalytic activity and leads to cellular aging and apoptosis. In experimental animal models, injection of GAPDH antagonists induces apoptosis and blocks Hep3B tumor progression, suggesting a therapeutic potential of targeting GAPDH in human hepatocellular carcinoma

Protein type: Carbohydrate Metabolism - glycolysis and gluconeogenesis; Oxidoreductase; EC 1.2.1.12

Chromosomal Location of Human Ortholog: 12p13

Cellular Component: microtubule cytoskeleton; nuclear membrane; membrane; intracellular membrane-bound organelle; perinuclear region of cytoplasm; cytoplasm; plasma membrane; lipid particle; ribonucleoprotein complex; cytosol; nucleus; vesicle

Molecular Function: identical protein binding; protein binding; microtubule binding; glyceraldehyde-3-phosphate dehydrogenase (phosphorylating) activity; NADP binding; NAD binding

Biological Process: neuron apoptosis; glycolysis; protein stabilization; negative regulation of translation; carbohydrate metabolic process; glucose metabolic process; pathogenesis; microtubule cytoskeleton organization and biogenesis; gluconeogenesis