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
Overview
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Product name
Anti-HPRT antibody
See all HPRT primary antibodies -
Description
Rabbit polyclonal to HPRT -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB MouseHuman
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Immunogen
Synthetic peptide corresponding to Human HPRT aa 200 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab24439) -
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
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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
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB MouseHumanAll applications RatChickenGerbilChinese hamsterApplication 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
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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
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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
see all -
Alternative names
- HGPRT antibody
- HGPRTase antibody
- HPRT 1 antibody
see all
Images
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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.
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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).
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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)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)
Protocols
Datasheets and documents
References (38)
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
Images
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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)
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).
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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)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)
