hh Human/Mouse/Rat GAPDH Antibody MAB5718: R&D Systems

Human/Mouse/Rat GAPDH Antibody

(2 citations)
(1 Review)
  
  • 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
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
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
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.
Detection of Human and Mouse GAPDH/G3PDH by Simple WesternTM. 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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Sample Type
  1. 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
  2. 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
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Mouse HRP-DAB Cell & Tissue Staining Kit

CTS002 14
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse IgG Antibody

WB AF007 4
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.

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Western Blot Human/Mouse/Rat GAPDH Antibody MAB5718
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 WB Human Anonymous 03/01/2017
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Western Blot Human/Mouse/Rat GAPDH Antibody MAB5718
Western Blot: Human/Mouse/Rat GAPDH Antibody [MAB5718]

Summary

ApplicationWestern Blot
Sample TestedHuman cell line
SpeciesHuman

Other Experimental Details

Other Experimental DetailsExpression of Human p75NGF in several cell lines, with GAPDH levels showing equal loading. Dilution: 1:10,000 in PBS with 5% BSA. Secondary Ab: 1:5,000 anti-mouse IgG.

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