Invitrogen

GAPDH Polyclonal Antibody

Product Image Gallery

GAPDH Antibody — Figure 5. G. intestinalis secreted cysteine proteases have effects on epithelial barrier proteins. A, B, C. Levels of tight junction and adherence junction proteins in Caco-2 whole cell extracts were examined after 24 h of recombinant cysteine proteases treatment (A, 14019; B, 16779; C, 16160), respectively, by Western blot analysis, with GAPDH used as a loading control. Barrier proteins were detected with a mouse or rabbit anti-barrier protein antibody and then incubated with a HR- conjugated mouse or rabbit antibody. D E, F. The localization of occludin in Caco-2 cells visualized by immunofluorescence microscopy. Caco-2 monolayers were incubated with 2.5 ug/ml of 14019 (D), 16779 (E), 16160 (F) in the absence or presence of E64 (10 uM) for 24 h, followed by fixation of PFA. Occludin was detected with a mouse anti-occludin antibody and then incubated with an Alexa Fluor 488-conjugated mouse antibody. Images were taken by a Zeiss Axioplan II Imaging fluorescence microscope and analyzed by ZEN 2.1 software. Bar = 50 um.

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).

Product Details

Tested Applications

Dilution

Western Blot (WB)

1:1000-1:5000

Published Applications

Western Blot (WB)

See 11 publications below

Product Specifications

Species Reactivity

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

Published species

Human, Mouse, Rat

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide: V(126) M G V N H E K Y(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; epididymis secretory sperm binding protein Li 162eP; G3PD; G3PDH; GAPD; GAPDH; Glyceraldehyde-3-Phosphate Dehydrogenase; Peptidyl-cysteine S-nitrosylase GAPDH

Gene Aliases: BARS-38; CDABP0047; G3PD; GAPD; GAPDH; HEL-S-162eP; OK/SW-cl.12

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

Entrez Gene ID: (Human) 2597, (Dog) 403755, (Mouse) 14433, (Rat) 24383

Molecular Function: dehydrogenase oxidoreductase