Recombinant 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
Related conjugates and formulations
Overview
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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/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5299 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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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 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 |
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WB |
1/10000 - 1/50000. Predicted molecular weight: 25 kDa.
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IP |
1/10 - 1/100.
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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 |
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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. |
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: 3251 Human
- Entrez Gene: 15452 Mouse
- Entrez Gene: 24465 Rat
- Omim: 308000 Human
- SwissProt: P00492 Human
- SwissProt: P00493 Mouse
- SwissProt: P27605 Rat
- Unigene: 412707 Human
see all -
Alternative names
- HGPRT antibody
- HGPRTase antibody
- HPRT 1 antibody
see all
Images
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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 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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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
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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. -
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.
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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 -
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (16)
ab109021 has been referenced in 16 publications.
- Svedlund Eriksson E et al. Castration of Male Mice Induces Metabolic Remodeling of the Heart. J Endocr Soc 6:bvac132 (2022). PubMed: 36249410
- Boldanova T et al. Supervised learning based on tumor imaging and biopsy transcriptomics predicts response of hepatocellular carcinoma to transarterial chemoembolization. Cell Rep Med 2:100444 (2021). PubMed: 34841291
- 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