Invitrogen

GAPDH Monoclonal Antibody (ZG003)

Product Image Gallery

GAPDH Antibody — Published figure using GAPDH monoclonal antibody (Product # 39-8600)

GAPDH Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of GAPDH was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with GAPDH Mouse Monoclonal Antibody (Product # 39-8600) at 1 µg/mL and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.

GAPDH Antibody in Western Blot (WB) — Western blot analysis of GAPDH was performed by loading 20 µg of A549 (lane1), A431 (lane2), U-87 MG (lane3), Mouse Lung (lane4), PC-3 (lane5), HeLa (Lane6) and K562 (lane7) lysate using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. GAPDH was detected at ~37 kDa using GAPDH Mouse Monoclonal Antibody (Product # 39-8600) at 0.5-1 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse IgG - HRP Secondary Antibody (Product # 62-6520) at 1:4000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

GAPDH Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of GAPDH showing staining in the cytoplasm and nucleus of paraffin-embedded human kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a GAPDH Mouse Monoclonal Antibody (ZG003) (Product # 39-8600) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Product Details

Tested Applications

Dilution

ELISA (ELISA)

Assay Dependent

Immunocytochemistry (ICC)

1-2 µg/mL

Immunofluorescence (IF)

1-2 µg/mL

Immunohistochemistry (Paraffin) (IHC (P))

1:20-1:200

Western Blot (WB)

0.5-2 µg/mL

Published Applications

Western Blot (WB)

See 32 publications below

Miscellaneous PubMed (MISC)

See 7 publications below

Immunofluorescence (IF)

See 3 publications below

Immunohistochemistry (IHC)

See 2 publications below

Product Specifications

Species Reactivity

Human, Mouse

Published species

Dog, Fish, Human, Mouse

Host / Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

ZG003

Immunogen

Recombinant full-length human GAPDH protein

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A

Storage buffer

PBS, pH 7.4

Contains

0.1% sodium azide

Storage conditions

-20°C

RRID

AB_2533438

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: aging-associated gene 9 protein; epididymis secretory sperm binding protein Li 162eP; G3PD; G3PDH; GAPD; GAPDH; Glyceraldehyde-3-Phosphate Dehydrogenase; Peptidyl-cysteine S-nitrosylase GAPDH

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

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

Entrez Gene ID: (Human) 2597, (Mouse) 14433

Molecular Function: dehydrogenase oxidoreductase