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Human/Mouse/Rat GAPDH Antibody

Catalog # AF5718 | R&D Systems, Inc. a Bio-Techne Brand
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AF5718
AF5718-SP

Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunocytochemistry, Immunohistochemistry, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Summary for Human/Mouse/Rat GAPDH Antibody

Immunogen

E. coli-derived recombinant human GAPDH/G3PDH
Met1-Ala150
Accession # P04406

Specificity

Detects human, mouse, and rat GAPDH/G3PDH in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human GAPDH-2 is observed. 

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat GAPDH Antibody

Detection of Human/Mouse/Rat GAPDH/G3PDH antibody by Western Blot.

Detection of Human/Mouse/Rat GAPDH/G3PDH by Western Blot.

Western blot shows lysates of mouse, human, and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat GAPDH/G3PDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GAPDH/G3PDH at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
GAPDH antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

GAPDH in HeLa Human Cell Line.

GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human/Mouse/Rat GAPDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
GAPDH/G3PDH antibody in Human Kidney by Immunohistochemistry (IHC-P).

GAPDH/G3PDH in Human Kidney.

GAPDH/G3PDH was detected in immersion fixed paraffin-embedded sections of normal human kidney using Goat Anti-Human/Mouse/Rat GAPDH/G3PDH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5718) at 8 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei of convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.This application has not been tested in mouse or rat samples.

Applications for Human/Mouse/Rat GAPDH Antibody

Application
Recommended Usage

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of normal human kidney and human prostate cancer tissue

Simple Western

10 µg/mL
Sample: Human brain (motor cortex) tissue and HepG2 human hepatocellular carcinoma cell line

Western Blot

1 µg/mL
Sample: Mouse, human, and rat brain tissue
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 9 reviews rated 4.9 using AF5718 in the following applications:

Published Applications

Read 24 publications using AF5718 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available:
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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: GAPDH

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a 36‑40 kDa member of the GAPDH family of enzymes. It is a widely expressed heterotetramer that is found in both the nucleus and cytoplasm. Although GAPDH was initially identified as a glycolytic enzyme that converted G3P into 1,3 diphosphoglycerate, it is now recognized to participate in no less than endocytosis, membrane fusion, vesicular secretory transport, DNA replication and repair, and apoptosis. Human GAPDH is 335 amino acids (aa) in length and contains two NAD binding sites (Asp35 and Asn316) with a catalytic region between aa 151‑155. GAPDH contains more than 19 posttranslational modifications, including methylation, deamidation and phosphorylation. One splice variant shows a 10 aa substitution for aa 319‑335. Over amino acid 1‑150, human GAPDH shares 92% aa identity with mouse GAPDH.

Long Name

Glyceraldehyde-3-phosphate Dehydrogenase

Alternate Names

G3PDH

Entrez Gene IDs

2597 (Human); 14433 (Mouse); 24383 (Rat)

Gene Symbol

GAPDH

UniProt

Product Documents for Human/Mouse/Rat GAPDH Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse/Rat GAPDH Antibody

For research use only

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Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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