Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-HPRT antibody [EPR5299] (ab109021)

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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

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

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. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

      see all

    • 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

    • Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
      Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)
      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
    • Immunoprecipitation - Anti-HPRT antibody [EPR5299] (ab109021)
      Immunoprecipitation - Anti-HPRT antibody [EPR5299] (ab109021)

      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 Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
      Western blot - Anti-HPRT antibody [EPR5299] (ab109021)

      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.

    • Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
      Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
      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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)

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

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    See all 7 Publications for this product

    Publishing research using ab109021? Please let us know so that we can cite the reference in this datasheet.

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    Immunocytochemistry/ Immunofluorescence abreview for Anti-HPRT antibody [EPR5299]

     Average
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 10 2015

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