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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-HPRT antibody [EPR5299] (ab109021)

Reviews (1) Submit a question References (12)
Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)
  • Immunoprecipitation - Anti-HPRT antibody [EPR5299] (ab109021)
  • Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
  • Western blot - Anti-HPRT antibody [EPR5299] (ab109021)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)
  • Anti-HPRT antibody [EPR5299] (ab109021)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5299] to HPRT
  • Suitable for: WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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. This information is proprietary to Abcam and/or its suppliers.

  • 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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR5299

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

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.
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.

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

References (12)

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

ab109021 has been referenced in 12 publications.

  • Mallampalli RK  et al. Cigarette smoke exposure enhances transforming acidic coiled-coil-containing protein 2 turnover and thereby promotes emphysema. JCI Insight 5:N/A (2020). PubMed: 31996486
  • Shimizu Y  et al. Paneth cell a-defensin misfolding correlates with dysbiosis and ileitis in Crohn's disease model mice. Life Sci Alliance 3:N/A (2020). PubMed: 32345659
  • Sabatino L  et al. Modification of cardiac thyroid hormone deiodinases expression in an ischemia/reperfusion rat model after T3 infusion. Mol Cell Biochem 475:205-214 (2020). PubMed: 32780210
  • Mardani I  et al. Plin2-deficiency reduces lipophagy and results in increased lipid accumulation in the heart. Sci Rep 9:6909 (2019). PubMed: 31061399
  • McGuinness D  et al. A molecular signature for delayed graft function. Aging Cell 17:e12825 (2018). PubMed: 30094915
View all Publications for this product

Customer reviews and Q&As

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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
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Verified customer

Submitted Aug 10 2015

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