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使用敲除细胞株进行验证重组RabMAb

重组Anti-HPRT抗体[EPR5299] (ab109021)

Reviews (1) Submit a question References (18)
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

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 APC Carrier Free PE

概述

  • 产品名称

    Anti-HPRT抗体[EPR5299]
    参阅全部 HPRT 一抗

  • 描述

    兔单克隆抗体[EPR5299] to HPRT

  • 宿主

    Rabbit

  • 特异性

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • 经测试应用

    适用于: WB, IP, IHC-Pmore details
    不适用于: Flow Cyt or ICC/IF

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

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

  • 阳性对照

    • 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

  • 常规说明

    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.

性能

  • 形式

    Liquid

  • 存放说明

    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.

  • 存储溶液

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

  • 纯度

    Protein A purified

  • 克隆

    单克隆

  • 克隆编号

    EPR5299

  • 同种型

    IgG

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab109021于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
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.
说明
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.
应用说明
Is unsuitable for Flow Cyt or ICC/IF.

靶标

  • 功能

    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.

  • 通路

    Purine metabolism; IMP biosynthesis via salvage pathway; IMP from hypoxanthine: step 1/1.

  • 疾病相关

    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.

  • 序列相似性

    Belongs to the purine/pyrimidine phosphoribosyltransferase family.

  • 细胞定位

    Cytoplasm.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • 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)

文献 (18)

发表研究结果有使用 ab109021？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab109021 被引用在 18 文献中.

  • Svedlund Eriksson E  et al. Castration of Male Mice Induces Metabolic Remodeling of the Heart. J Endocr Soc 6:bvac132 (2022). PubMed: 36249410
  • Kretzmer C  et al. De novo assembly and annotation of the CHOZN® GS-/- genome supports high-throughput genome-scale screening. Biotechnol Bioeng 119:3632-3646 (2022). PubMed: 36073082
  • 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
  • Nowaczyk M  et al. Addition of Popular Exogenous Antioxidant Agent, PBN, to Culture Media May Be an Important Step to Optimization of Myogenic Stem/Progenitor Cell Preparation Protocol. Antioxidants (Basel) 10:N/A (2021). PubMed: 34203726
  • 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
View all Publications for this product

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Immunocytochemistry/ Immunofluorescence abreview for Anti-HPRT antibody [EPR5299]

 Average
Abreviews
This product is known to not work in this application or species.
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Aug 10 2015

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