Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-HPRT antibody [EPR5298] (ab133242)

Overview

  • Product name

    Anti-HPRT antibody [EPR5298]
    See all HPRT primary antibodies
  • Description

    Rabbit monoclonal [EPR5298] to HPRT
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues in human HPRT (P00492).

  • Positive control

    • WB: Wild-type HAP1, HeLa, HepG2, A431 and MCF7 cell lysates.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133242 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 1/1000 - 1/10000. Predicted molecular weight: 25 kDa.
IP Use at an assay dependent dilution.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • 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

    • 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 - ab133242 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab133242 was shown to specifically react with HPRT1 in wild-type HAP1 cells. No band was observed when HPRT1 knockout samples were used. Wild-type and HPRT1 knockout samples were subjected to SDS-PAGE.  ab133242 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 and 1/10,000 respectively. 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 [EPR5298] (ab133242) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 25 kDa

    References

    ab133242 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Placenta)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Placenta
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Jeremiah Gaudet

    Verified customer

    Submitted Aug 03 2017

    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Placenta)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Placenta
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Jeremiah Gaudet

    Verified customer

    Submitted Aug 03 2017

    Answer

    The AbTrial is not applicable to tests of ab133242 for staining western blots of mouse samples, as this combination (western blotting and mouse) has already been tested. Our Abpromise guarantee will apply, though, as it covers western blotting of mouse samples with ab133242. For details of our guarantee, please see the page at www.abcam.com/abpromise .

    Your plan to stain the same blot with two different antibodies will not be difficult. Most people attempt this by cutting the membrane in half. The two proteins you are trying to detect are 25 kDa (HPRT) and 53-60 kDa (5HT2A receptor). So, if you cut the membrane at about 40 kDa, using pre-stained molecular weight markers to guide you, you can stain each half separately, one half for each antibody.

    If you do not cut the membrane, and stain it with both antibodies, then you will not know which bands were produced by which antibody.

    Read More

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