Invitrogen

GAPDH Polyclonal Antibody

9 Published Figures

27 References

View all (79) GAPDH antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

GAPDH Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), PC-3 (Lane 4) and tissue extract (30 µg lysate) of Ms Brain (Lane 5). The blot was probed with Anti-GAPDH Polyclonal Antibody (Product # PA1-987, 1:1,000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 37 kDa band corresponding to GAPDH was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

GAPDH Antibody — FIG 1 Caspase-3 is required for deoxyadenosine-induced killing of macrophages. (A) U937-derived macrophages (MPhi) were treated with (+) or without (-) dAdo, and cell lysates were analyzed for caspase-3 activity using a colorimetric assay. As controls, lysates were treated with the caspase-3 inhibitor Ac-DEVD-CHO (+ Inhib.). (B) Survival of MPhi exposed to dAdo and increasing concentrations of Z-DEVD-FMK (0 to 5 muM), an inhibitor of caspase-3. (C) Diagram illustrating the position of CASP3 on chromosome 4 and exons 1 to 8 of CASP3 mRNA. Sequencing results for mutated exon 5 alleles (red box) cloned from CASP3 -/- cells are shown. (D) Immunoblotting of lysates from wild-type (WT) MPhi and their CASP3 -/- and complemented CASP3 -/- variants (+ CASP3 WT ) with caspase-3 and GAPDH-specific antibodies (alpha-CASP3 and alpha-GAPDH, respectively). GAPDH was used as a loading control. Numbers to the left of blots indicate the migration of molecular weight markers in kilodaltons. (E and F) Survival of MPhi (black bars) and their SLC29A1 -/- (blue bars), CASP3 -/- (red bars), and complemented CASP3 -/- variants (+ CASP3 WT , white bars) after treatment with dAdo (E) or after treatment with culture medium (RPMI 1640) that had been conditioned by incubation with either wild-type S. aureus Newman or its adsA mutant in the presence of host DNA, as indicated by + and - signs (F). All samples received adenosine deaminase inhibitor (50 muM dCF). Data are the mean (+- standard deviation [SD])

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1000-1:5000

View 27 publications 27 publications

Product Specifications

Species

Dog, Human, Mouse, Non-human primate, Rat

Published species

Artificial Control, Human, Mouse, Rat

Expression System

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide: A(126) DAPMFVMGVN HEKY(140)

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS with 1mg/mL BSA

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2107311

Product Specific Information

PA1-987 detects rat GAPDH. Shares 100% sequence homology with human and mouse.

PA1-987 has been used successfully in Western blotting. By Western blot this antibody specifically detects a ~36 kDa protein representing GAPDH.

PA1-987 immunizing peptide corresponds to amino acid residues 126-140 from human GAPDH.

Target Information

GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) is a catalytic enzyme commonly known to be involved in glycolysis. GAPDH exists as a tetramer of identical 37-kDa subunits and catalyzes the reversible reduction of 1,3-bisphosphoglycerate to glyceraldehyde 3-phosphophate in the presence of NADPH. Apart from playing a key role in glycolysis, GAPDH is ubiquitously expressed and displays other activities unrelated to its glycolytic function. GAPDH is reported to be involved in the processes of DNA replication, DNA repair, nuclear RNA export, membrane fusion and microtubule bundling. Studies provide evidence of GAPDH playing an essential part in gene expression observed in apoptosis and as part of the cellular phenotype of age-related neurodegenerative diseases. Further, GAPDH is involved in other cellular processes ranging from membrane fusion, and neuronal apoptosis in cancer. GAPDH is reported to bind to a variety of other proteins, including the amyloid precursor protein, mutations in which cause some forms of Alzheimer's disease (AD), and the polyglutamine tracts of Huntingtin, the protein product aberrant forms of which are causative of Huntington's disease. Associations between GAPDH, actin and tubulin have also be reported. Since GAPDH is expressed at high levels in most tissues, it is useful as protein loading control in Western Blot analysis.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: 38 kDa BFA-dependent ADP-ribosylation substrate; aging-associated gene 9 protein; BARS-38; EC 1.2.1.12; epididymis secretory protein Li 278; epididymis secretory sperm binding protein Li 162eP; GAPDH; GAPDH-2; Glyceraldehyde-3-phosphate dehydrogenase; glyceraldehyde-3-phosphate dehydrogenase type 2; Glyceraldehyde-3-phosphate dehydrogenase, testis-specific; HGNC:4141; MGC88685; Peptidyl-cysteine S-nitrosylase GAPDH; Spermatogenic cell-specific glyceraldehyde 3-phosphate dehydrogenase 2; Spermatogenic glyceraldehyde-3-phosphate dehydrogenase

Gene Aliases: BARS-38; CDABP0047; G3PD; GAPD; Gapd-s; GAPD2; GAPDH; GAPDH-2; GAPDH2; GAPDHS; GAPDS; HEL-S-162eP; HEL-S-278; HSD-35; HSD35; OK/SW-cl.12

UniProt ID: (Human) P04406, (Human) O14556, (Mouse) P16858, (Rat) P04797, (Rat) Q9ESV6, (Mouse) Q64467

Entrez Gene ID: (Human) 2597, (Dog) 403755, (Human) 26330, (Dog) 476483, (Mouse) 14433, (Rat) 24383, (Rat) 66020, (Mouse) 14447

Molecular Function: dehydrogenase oxidoreductase