Overview

  • Product nameAnti-GAPDH antibodySee all GAPDH primary antibodies ...
  • Description
    Rabbit polyclonal to GAPDH
  • Tested applicationsICC/IF, IHC-P, WB more details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Guinea pig, Turkey, Pig, Xenopus laevis, Chimpanzee, Reptiles, Rhesus monkey, Gorilla
  • Immunogen

    Synthetic peptide, corresponding to a region between residues 250-300 of Human GAPDH. (NP_002037.2).

  • Positive control
    • Whole cell lysate from HeLa and mouse NIH3T3 cells. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal skeletal muscle. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.

Properties

Applications

Our Abpromise guarantee covers the use of ab70699 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml.
WB 1/2000 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 36 kDa).
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionHas both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate.
    • PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
    • Sequence similaritiesBelongs to the glyceraldehyde-3-phosphate dehydrogenase family.
    • Post-translational
      modifications
      S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
      ISGylated.
    • Cellular localizationCytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
    • Target information above from: UniProt accession P04406 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links
    • Alternative names
      • 38 kDa BFA-dependent ADP-ribosylation substrate antibody
      • aging associated gene 9 protein antibody
      • Aging-associated gene 9 protein antibody
      • BARS-38 antibody
      • cb609 antibody
      • EC 1.2.1.12 antibody
      • G3P_HUMAN antibody
      • G3PD antibody
      • G3PDH antibody
      • GAPD antibody
      • GAPDH antibody
      • GAPDH antibody
      • Glyceraldehyde 3 phosphate dehydrogenase antibody
      • Glyceraldehyde-3-phosphate dehydrogenase antibody
      • KNC-NDS6 antibody
      • MGC102544 antibody
      • MGC102546 antibody
      • MGC103190 antibody
      • MGC103191 antibody
      • MGC105239 antibody
      • MGC127711 antibody
      • MGC88685 antibody
      • OCAS, p38 component antibody
      • OCT1 coactivator in S phase, 38-KD component antibody
      • peptidyl cysteine S nitrosylase GAPDH antibody
      • Peptidyl-cysteine S-nitrosylase GAPDH antibody
      • wu:fb33a10 antibody
      see all

    Anti-GAPDH antibody images

    • ab70699 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab70699 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • All lanes : Anti-GAPDH antibody (ab70699) at 0.04 µg/ml

      Lane 1 : Whole cell lysate from HeLa cells at 50 µg
      Lane 2 : Whole cell lysate from HeLa cells at 15 µg
      Lane 3 : Whole cell lysate from HeLa cells at 5 µg
      Lane 4 : Whole cell lysate from NIH 3T3 cells at 50 µg


      Predicted band size : 36 kDa
      Observed band size : 38 kDa (why is the actual band size different from the predicted?)


      Exposure time : 10 seconds
    • IHC image of GAPDH staining in Human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70699, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    References for Anti-GAPDH antibody (ab70699)

    This product has been referenced in:
    • Charron G  et al. Prenylome profiling reveals S-farnesylation is crucial for membrane targeting and antiviral activity of ZAP long-isoform. Proc Natl Acad Sci U S A 110:11085-90 (2013). Read more (PubMed: 23776219) »
    • Yount JS  et al. S-Palmitoylation and Ubiquitination Differentially Regulate Interferon-induced Transmembrane Protein 3 (IFITM3)-mediated Resistance to Influenza Virus. J Biol Chem 287:19631-41 (2012). WB ; Human . Read more (PubMed: 22511783) »

    See all 2 Publications for this product

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