Human/Mouse/Rat GAPDH Antibody

Name: Human/Mouse/Rat GAPDH Antibody
Brand: R&D Systems
Rating: 5 (1 reviews)

(2 citations)

(1 Review)

Western Blot

Immunocytochemistry

Immunohistochemistry

Simple Western

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse and rat GAPDH/G3PDH in Western blots. In direct ELISAs, 100% cross-reactivity with recombinant mouse GAPDH/G3PDH and no cross-reactivity with recombinant human GAPDH-2 is observed.
Source
Monoclonal Mouse IgG₁ Clone # 686613
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human GAPDH/G3PDH
Met1-Ala150
Accession # P04406
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended
Concentration

Sample

Western Blot

0.05 µg/mL

See below
Simple Western

0.25 µg/mL

See below
Immunohistochemistry

8-25 µg/mL

See below
Immunocytochemistry

8-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot click image to view larger	Detection of Human, Mouse, and Rat GAPDH/G3PDH by Western Blot. Western blot shows lysates of human brain tissue, mouse brain tissue, and rat brain tissue. PVDF Membrane was probed with 0.05 µg/mL of Mouse Anti-Human/Mouse/Rat GAPDH/G3PDH Monoclonal Antibody (Catalog # MAB5718) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for GAPDH/G3PDH at approximately 39 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Immunocytochemistry click image to view larger	GAPDH in HeLa Human Cell Line. GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat GAPDH Monoclonal Antibody (Catalog # MAB5718) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry click image to view larger	GAPDH/G3PDH in Human Kidney. GAPDH/G3PDH was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-Human/Mouse/Rat GAPDH/G3PDH Monoclonal Antibody (Catalog # MAB5718) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Simple Western click image to view larger	Detection of Human and Mouse GAPDH/G3PDH by Simple Western^TM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and C2C12 mouse myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for GAPDH/G3PDH at approximately 41 kDa (as indicated) using 0.25 µg/mL of Mouse Anti-Human/Mouse/Rat GAPDH/G3PDH Monoclonal Antibody (Catalog # MAB5718). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
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

GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) 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. There are two NAD binding sites (Asp35 and Asn316) with a catalytic region between aa 151-155. GAPDH contains no fewer that 19 posttranslational modifications, including methylation, deamidation and phosphorylation. One splice variant shows a 10 aa substitution for aa 319‑335. Over
aa 1‑150, human GAPDH shares 92% aa identity with mouse GAPDH.

Long Name:
Glyceraldehyde-3-phosphate Dehydrogenase
Entrez Gene IDs:
2597 (Human); 14433 (Mouse); 24383 (Rat)
Alternate Names:
aging-associated gene 9 protein; EC 1.2.1; EC 1.2.1.12; EC 2.6.99.-; G3PD; G3PDH; GAPDH; GAPDPeptidyl-cysteine S-nitrosylase GAPDH; glyceraldehyde 3-phosphate dehydrogenase; glyceraldehyde-3-phosphate dehydrogenase; MGC88685

Related Research Areas

Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Species	Applications	Sample Type
The transcription factor MITF is a critical regulator of GPNMB expression in dendritic cells. Authors: Gutknecht M, Geiger J, Joas S, Dorfel D, Salih H, Muller M, Grunebach F, Rittig S Cell Commun Signal, 2015;13(0):19. Species: Human Sample Type: Cell Lysates Application: WB Normal leptin expression, lower adipogenic ability, decreased leptin receptor and hyposensitivity to Leptin in Adolescent Idiopathic Scoliosis. Authors: Liang G, Gao W, Liang A PLoS ONE, 2012;7(5):e36648. Species: Human Sample Type: Cell Lysates Application: WB

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Cell and Tissue Staining Kits

Description Application Cat# Citations Images

Anti-Mouse HRP-DAB Cell & Tissue Staining Kit

CTS002 14

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Human/Mouse/Rat GAPDH Antibody

Anti-Mouse HRP-DAB Cell & Tissue Staining Kit

Antigen Retrieval Reagent-Universal

Antigen Retrieval Reagent-Basic

Mounting Medium

Antigen Retrieval Reagent Sampler (50 mL each)

Antigen Retrieval Reagent-Acidic

NorthernLights Guard Mounting Media

VisUCyte Antigen Retrieval Reagent-Acidic

VisUCyte Antigen Retrieval Reagent-Basic

VisUCyte Antigen Retrieval Reagent-Universal

Mouse IgG1 Isotype Control

Mouse IgG1 Isotype Control

Mouse IgG HRP-conjugated Antibody

Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Mouse IgG HRP-conjugated Antibody

Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Donkey Anti-Mouse IgG Biotinylated Antibody

Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Mouse IgG Antibody

Goat Anti-Mouse IgG Biotinylated Antibody

Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

Mouse IgG VisUCyte HRP Polymer Antibody

Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

Donkey Anti-Mouse IgG (H+L) Affinity Purified PAb, X Absorb

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