Recombinant Anti-HPRT antibody [EPR5299] (ab109021)
![Western blot - Anti-HPRT antibody [EPR5299] (ab109021)](https://www.abcam.com/ps/products/109/ab109021/Images/ab109021-314278-anti-hprt-antibody-epr5299-western-blot.jpg "Western blot - 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
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 & Species
See all applications and species dataApplication Species IHC-P HumanIP RatWB MouseRatHuman
-
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
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
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| IHC-P |
Human
|
| IP |
Rat
|
| WB |
Mouse
Rat
Human
|
| 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. |
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
- 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
-
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 kDaBlocking and diluting buffer: 5% NFDM/TBST.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)](https://www.abcam.com/ps/products/109/ab109021/Images/ab109021-314311-anti-hprt-antibody-epr5299-immunohistochemistry.jpg)
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)](https://www.abcam.com/ps/products/109/ab109021/Images/ab109021-314346-anti-hprt-antibody-epr5299-immunoprecipitation.jpg)
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)](https://www.abcam.com/ps/products/109/ab109021/Images/ab109021-270766-anti-hprt-antibody-epr5299-western-blot.jpg)
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)](https://www.abcam.com/ps/products/109/ab109021/Images/ab109021-121599-anti-hprt-antibody-epr5299-western-blot.jpg)
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)](https://www.abcam.com/ps/products/109/ab109021/Images/ab109021-121598-anti-hprt-antibody-epr5299-immunohistochemistry.jpg)
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)](https://www.abcam.com/ps/products/109/ab109021/Images/ab109021-7-benefits-of-recombinant-antibodies.png)
Anti-HPRT antibody [EPR5299] (ab109021)
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (10)
ab109021 has been referenced in 10 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
- 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
- Wang X et al. MicroRNA-145 Aggravates Hypoxia-Induced Injury by Targeting Rac1 in H9c2 Cells. Cell Physiol Biochem 43:1974-1986 (2017). PubMed: 29055960
- McLaughlin D et al. Signalling mechanisms underlying doxorubicin and Nox2 NADPH oxidase-induced cardiomyopathy: involvement of mitofusin-2. Br J Pharmacol 174:3677-3695 (2017). PubMed: 28261787
- Dominici C et al. Floor-plate-derived netrin-1 is dispensable for commissural axon guidance. Nature 545:350-354 (2017). PubMed: 28445456
- Koch Y et al. Aggregated a-Synuclein Increases SOD1 Oligomerization in a Mouse Model of Amyotrophic Lateral Sclerosis. Am J Pathol 186:2152-2161 (2016). PubMed: 27322773
- Moscato S et al. Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study. J Funct Biomater 6:16-32 (2015). PubMed: 25590431
- Andersson L et al. Rip2 modifies VEGF-induced signalling and vascular permeability in myocardial ischaemia. Cardiovasc Res 107:478-86 (2015). PubMed: 26130752