Human/Mouse/Rat GAPDH Antibody

  (3 citations)
(2 Reviews)
    
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Product Details
Citations (3)
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  • 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
Product Datasheets

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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.
    Reconstitution Buffer Available
  • 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.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Characterization of the Giardia intestinalis secretome during interaction with human intestinal epithelial cells: The impact on host cells
    Authors: SY Ma'ayeh, J Liu, D Peirasmaki, K Hörnaeus, S Bergström, M Grabherr, J Bergquist, SG Svärd
    PLoS Negl Trop Dis, 2017;11(12):e0006120.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. 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
  3. 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

FAQs

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Isotype Controls
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Average Rating: 5 (Based on 2 reviews)

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Read what people are saying who have used Human/Mouse/Rat GAPDH Antibody.

We have 2 reviews tested in 2 species: Human, Human,Mouse.
We have 2 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Excellent
 WB Human,Mouse Anonymous 07/03/2018
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Summary

Application Western Blot
Sample Tested Cancer cell lysates,Xenograft Lysates
Species Human,Mouse
Western Blot Human/Mouse/Rat GAPDH Antibody MAB5718 Array MAB5718
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Excellent
 WB Human Anonymous 03/01/2017
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Image Details
Western Blot Human/Mouse/Rat GAPDH Antibody MAB5718
Western Blot: Human/Mouse/Rat GAPDH Antibody [MAB5718].

Summary

Application Western Blot
Sample Tested Human cell line
Species Human

Other Experimental Details

Other Experimental Details Expression 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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