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    hprt-antibody-ab10479.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA / Nucleotides
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Validated using a knockout cell line

Anti-HPRT antibody (ab10479)

  • Datasheet
  • SDS
Reviews (4) Submit a question References (38)

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Western blot - Anti-HPRT antibody (ab10479)
  • Immunocytochemistry/ Immunofluorescence - Anti-HPRT antibody (ab10479)
  • Western blot - Anti-HPRT antibody (ab10479)
  • Western blot - Anti-HPRT antibody (ab10479)
  • Western blot - Anti-HPRT antibody (ab10479)

Key features and details

  • Rabbit polyclonal to HPRT
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-HPRT antibody
    See all HPRT primary antibodies
  • Description

    Rabbit polyclonal to HPRT
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    WB
    Mouse
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human HPRT aa 200 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab24439)

  • Positive control

    • Recombinant Human HPRT protein (ab117153) can be used as a positive control in WB. This antibody gave a positive control in the following human whole cell lysates: HeLa, A431, MCF-7, HEK 293 whole cell lysate This antibody gave a positive control in the following mouse lysates: NIH 3T3, MEF1 whole cell lysate, mouse brain tissue lysate

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA / Nucleotides
    • Cell Biology
    • Other Antibodies
    • Other Antibodies

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human HPRT protein (Active) (ab222951)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab10479 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
WB
Mouse
Human
All applications
Rat
Chicken
Gerbil
Chinese hamster
Application Abreviews Notes
WB (4)
1/500 - 1/1000. Predicted molecular weight: 24 kDa.
ICC/IF
Use a concentration of 5 µg/ml.
Notes
WB
1/500 - 1/1000. Predicted molecular weight: 24 kDa.
ICC/IF
Use a concentration of 5 µg/ml.

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.
  • Target information above from: UniProt accession P00492 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 395653 Chicken
    • Entrez Gene: 3251 Human
    • Entrez Gene: 15452 Mouse
    • Entrez Gene: 24465 Rat
    • Omim: 308000 Human
    • SwissProt: Q9W719 Chicken
    • SwissProt: P00492 Human
    • SwissProt: P00493 Mouse
    • SwissProt: P27605 Rat
    • Unigene: 412707 Human
    • Unigene: 299381 Mouse
    • Unigene: 47 Rat
    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 (ab10479)
    Western blot - Anti-HPRT antibody (ab10479)

    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 - ab10479 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab10479 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.  Ab10479 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 dilution 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-HPRT antibody (ab10479)
    Immunocytochemistry/ Immunofluorescence - Anti-HPRT antibody (ab10479)

    ICC/IF image of ab10479 stained human HeLa cells. The cells were methanol fixed ( 5 min) and incubated with the antibody (ab10479, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).

  • Western blot - Anti-HPRT antibody (ab10479)
    Western blot - Anti-HPRT antibody (ab10479)
    All lanes : Anti-HPRT antibody (ab10479) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : A-431 whole cell lysate (ab7909)
    Lane 3 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK-293 whole cell lysate (ab7902)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 24 kDa

  • Western blot - Anti-HPRT antibody (ab10479)
    Western blot - Anti-HPRT antibody (ab10479)
    All lanes : Anti-HPRT antibody (ab10479) at 1 µg/ml

    Lane 1 : NIH/3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 24 kDa
    Observed band size: 24 kDa
    Additional bands at: 28 kDa (possible glycosylated form)

  • Western blot - Anti-HPRT antibody (ab10479)
    Western blot - Anti-HPRT antibody (ab10479)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (38)

Publishing research using ab10479? Please let us know so that we can cite the reference in this datasheet.

ab10479 has been referenced in 38 publications.

  • Chan DY  et al. Multigene human artificial chromosome vector delivery with herpes simplex virus 1 amplicons. Exp Cell Res 388:111840 (2020). PubMed: 31930965
  • Kofuji S  et al. IMP dehydrogenase-2 drives aberrant nucleolar activity and promotes tumorigenesis in glioblastoma. Nat Cell Biol 21:1003-1014 (2019). PubMed: 31371825
  • Köhler JC  et al. Early-Life Adversity Induces Epigenetically Regulated Changes in Hippocampal Dopaminergic Molecular Pathways. Mol Neurobiol 56:3616-3625 (2019). PubMed: 30173406
  • Fernandez-Mosquera L  et al. Mitochondrial respiratory chain deficiency inhibits lysosomal hydrolysis. Autophagy 15:1572-1591 (2019). PubMed: 30917721
  • Spychala A & Rüther U FTO affects hippocampal function by regulation of BDNF processing. PLoS One 14:e0211937 (2019). PubMed: 30730976
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-HPRT antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (THP1 Cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
THP1 Cells
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5%
Read More

Abcam user community

Verified customer

Submitted Jul 27 2018

Western blot

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin)
Sample
Mouse Cell lysate - whole cell (beta cells)
Specification
beta cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Jun 05 2013

Western blot abreview for Anti-HPRT antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Hippocampus)
Loading amount
5 µg
Specification
Hippocampus
Gel Running Conditions
Reduced Denaturing (12% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Sep 02 2010

Western blot abreview for Anti-HPRT antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse striatum and liver)
Loading amount
20 µg
Specification
Mouse striatum and liver
Gel Running Conditions
Reduced Denaturing (4-12% TG)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Dr. Laura Ling

Verified customer

Submitted Jul 31 2008

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