Validated using a knockout cell line
Recombinant
RabMAb

Anti-HPRT antibody [EPR5299] (ab109021)

Overview

  • Product name
    Anti-HPRT antibody [EPR5299]
    See all HPRT primary antibodies
  • Description
    Rabbit monoclonal [EPR5299] to HPRT
  • Host species
    Rabbit
  • Specificity
    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
  • Tested applications
    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human HPRT aa 200-300. The exact sequence is proprietary.

  • Positive control
    • WB: HeLa HepG2, A549, and MCF7 cell lysates, Mouse brain and Rat brain tissue lysates IHC-P:Human lung and colon cancer IP: Rat brain lysate
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109021 in the following tested applications.

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

Application Abreviews Notes
WB 1/10000 - 1/50000. Predicted molecular weight: 25 kDa.
IP 1/10 - 1/100.
IHC-P 1/150.

Heat up to 98 °C, below boiling, and then let cool for 10-20 min. See IHC antigen retrieval protocols.

For unpurified use at 1/50 - 1/100

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

 

  • Application notes
    Is unsuitable for Flow Cyt or ICC.
  • Target

    • Function
      Converts guanine to guanosine monophosphate, and hypoxanthine to inosine monophosphate. Transfers the 5-phosphoribosyl group from 5-phosphoribosylpyrophosphate onto the purine. Plays a central role in the generation of purine nucleotides through the purine salvage pathway.
    • Pathway
      Purine metabolism; IMP biosynthesis via salvage pathway; IMP from hypoxanthine: step 1/1.
    • Involvement in disease
      Defects in HPRT1 are the cause of Lesch-Nyhan syndrome (LNS) [MIM:300322]. LNS is characterized by complete lack of enzymatic activity that results in hyperuricemia, choreoathetosis, mental retardation, and compulsive self-mutilation.
      Defects in HPRT1 are the cause of gout HPRT-related (GOUT-HPRT) [MIM:300323]; also known as HPRT-related gout or Kelley-Seegmiller syndrome. Gout is characterized by partial enzyme activity and hyperuricemia.
    • Sequence similarities
      Belongs to the purine/pyrimidine phosphoribosyltransferase family.
    • Cellular localization
      Cytoplasm.
    • Information by UniProt
    • Database links
    • Alternative names
      • HGPRT antibody
      • HGPRTase antibody
      • HPRT 1 antibody
      • HPRT_HUMAN antibody
      • HPRT1 antibody
      • Hypoxanthine guanine phosphoribosyltransferase antibody
      • Hypoxanthine phosphoribosyltransferase 1 (Lesch Nyhan syndrome) antibody
      • Hypoxanthine phosphoribosyltransferase 1 antibody
      • Hypoxanthine-guanine phosphoribosyltransferase antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling HPRT with Purified ab109021 at 1:150 dilution (1.19 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
    • ab109021 (purified) at 1:30 dilution (2µg) immunoprecipitating HPRT in Rat brain lysate.
      Lane 1 (input): Rat brain lysate 10µg
      Lane 2 (+): ab109021 & Rat brain lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109021 in Rat brain lysate
      For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.
    • All lanes : Anti-HPRT antibody [EPR5299] (ab109021) at 0.02 µg/ml (purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : Mouse brain lysates
      Lane 3 : Rat brain lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.02 µg/ml

      Predicted band size: 25 kDa



      Blocking and diluting buffer: 5% NFDM/TBST.

    • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: HPRT1 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: A431 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab109021 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Unpurified ab109021 was shown to specifically react with HPRT1 in wild-type HAP1 cells. No band was observed when HPRT1 knockout samples were examined. Wild-type and HPRT1 knockout samples were subjected to SDS-PAGE. ab109021 and ab8245 (Mouse anti GAPDH loading control) were both diluted at 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-HPRT antibody [EPR5299] (ab109021) at 1/10000 dilution (Unpurified)

      Lane 1 : HeLa cell lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : A549 cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 25 kDa

    • Unpurified ab109021 at 1/50 dilution staining HPRT in Human colon by Immunohistochemistry, Paraffin-embedded tissue.

    References

    This product has been referenced in:
    • Moscato S  et al. Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study. J Funct Biomater 6:16-32 (2015). Read more (PubMed: 25590431) »
    • Andersson L  et al. Rip2 modifies VEGF-induced signalling and vascular permeability in myocardial ischaemia. Cardiovasc Res 107:478-86 (2015). Read more (PubMed: 26130752) »

    See all 2 Publications for this product

    Customer reviews and Q&As

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (A549)
    Permeabilization
    Yes - Triton X, 0.025%
    Specification
    A549
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Methanol
    Username

    Abcam user community

    Verified customer

    Submitted Aug 10 2015

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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