使用敲除细胞株进行验证RabMAb

Anti-Chk2抗体[EPR4325] (ab109413)

概述

  • 产品名称Anti-Chk2抗体[EPR4325]
    参阅全部 Chk2 一抗
  • 描述
    兔单克隆抗体[EPR4325] to Chk2
  • 经测试应用适用于: ICC/IF, WB, IP, IHC-P, ICCmore details
    不适用于: Flow Cyt
  • 种属反应性
    与反应: Human
  • 免疫原

    Recombinant fragment corresponding to Human Chk2 aa 1-200.

  • 阳性对照
    • HeLa treated with Gamma Irradiation, HeLa, HT-29, and 293T cell lysates; Human colon tissue; Human spleen tissue ICC/IF: HAP1 wild-type cells and HAP1-Chk2 knockout cells
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative versions available:

    Anti-Chk2 antibody (Alexa Fluor® 488) [EPR4325] (ab200762)
    Anti-Chk2 antibody (HRP) [EPR4325] (ab200767)

性能

应用

Our Abpromise guarantee covers the use of ab109413 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF 1/250.
WB 1/50000 - 1/200000. Detects a band of approximately 62 kDa (predicted molecular weight: 61 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. antigen retrieval is recommended.
ICC 1/100 - 1/250.
  • 应用说明Is unsuitable for Flow Cyt.
  • 靶标

    • 功能Regulates cell cycle checkpoints and apoptosis in response to DNA damage, particularly to DNA double-strand breaks. Inhibits CDC25C phosphatase by phosphorylation on 'Ser-216', preventing the entry into mitosis. May also play a role in meiosis. Regulates the TP53 tumor suppressor through phosphorylation at 'Thr-18' and 'Ser-20'.
    • 组织特异性High expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.
    • 疾病相关Defects in CHEK2 are associated with Li-Fraumeni syndrome 2 (LFS2) [MIM:609265]; a highly penetrant familial cancer phenotype usually associated with inherited mutations in p53/TP53.
      Defects in CHEK2 may be a cause of susceptibility to prostate cancer (PC) [MIM:176807]. It is a malignancy originating in tissues of the prostate. Most prostate cancers are adenocarcinomas that develop in the acini of the prostatic ducts. Other rare histopathologic types of prostate cancer that occur in approximately 5% of patients include small cell carcinoma, mucinous carcinoma, prostatic ductal carcinoma, transitional cell carcinoma, squamous cell carcinoma, basal cell carcinoma, adenoid cystic carcinoma (basaloid), signet-ring cell carcinoma and neuroendocrine carcinoma.
      Defects in CHEK2 are found in some patients with osteogenic sarcoma (OSRC) [MIM:259500].
    • 序列相似性Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHK2 subfamily.
      Contains 1 FHA domain.
      Contains 1 protein kinase domain.
    • 翻译后修饰Phosphorylated by PLK4.
    • 细胞定位Nucleus; Nucleus. Isoform 10 is present throughout the cell and Nucleus > PML body. Nucleus > nucleoplasm. Recruited into PML bodies together with TP53.
    • Information by UniProt
    • 数据库链接
    • 别名
      • CDS 1 antibody
      • Cds1 antibody
      • Cds1 homolog antibody
      • Checkpoint kinase 2 antibody
      • Checkpoint like protein CHK2 antibody
      • CHEK 2 antibody
      • Chek2 antibody
      • Chk 2 antibody
      • CHK2 checkpoint homolog (S. pombe) antibody
      • CHK2 checkpoint homolog antibody
      • CHK2_HUMAN antibody
      • hCds1 antibody
      • HuCds 1 antibody
      • LFS 2 antibody
      • LFS2 antibody
      • PP1425 antibody
      • RAD 53 antibody
      • RAD53 antibody
      • Rad53 homolog antibody
      • Serine/threonine protein kinase Chk2 antibody
      • Serine/threonine-protein kinase Chk2 antibody
      see all

    Anti-Chk2 antibody [EPR4325] 图像



    • Predicted band size : 61 kDa

      Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 6 and 10: Chk2 knockout HAP1 cell lysate (20 µg)
      Lanes 3, 7 and 11: HeLa cell lysate (20 µg)
      Lanes 4, 8 and 12: HEK293 cell lysate (20 µg)
      Lanes 1, 2, 3 and 4: Green signal from target -  ab109413 observed at 62 kDa
      Lanes 5, 6, 7 and 8: Red signal from loading control - ab8245 observed at 37 kDa
      Lanes 9, 10, 11 and 12: Merged (red and green) signal
      ab109413 was shown to specifically react with Chk2 when Chk2 knockout samples were used. Wild-type and Chk2 knockout samples were subjected to SDS-PAGE. ab109413 and ab8245 (loading control to GAPDH) were diluted 1/50 000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • ab109413 staining Chk2 in wild-type HAP1 cells (top panel) and Chk2 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109413 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • Immunohistochemical analysis of paraffin-embedded Human colon tissue using 1/100 ab109413
    • All lanes : Anti-Chk2 antibody [EPR4325] (ab109413) at 1/50000 dilution

      Lane 1 : HeLa treated with Gamma Irradiation
      Lane 2 : HeLa cell lysate
      Lane 3 : HT29 cell lysate
      Lane 4 : 293T cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 61 kDa
      Observed band size : 62 kDa (why is the actual band size different from the predicted?)
    • Immunohistochemical analysis of paraffin-embedded Human spleen tissue using 1/100 ab109413
    • ab109413 (1/500) staining Chk2 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5%Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please see Abreview.

      See Abreview

    Anti-Chk2 antibody [EPR4325] (ab109413)参考文献

    This product has been referenced in:
    • Li WF  et al. WISP-1 contributes to fractionated irradiation-induced radioresistance in esophageal carcinoma cell lines and mice. PLoS One 9:e94751 (2014). Human . Read more (PubMed: 24728101) »
    • Sowd GA  et al. SV40 utilizes ATM kinase activity to prevent non-homologous end joining of broken viral DNA replication products. PLoS Pathog 10:e1004536 (2014). Read more (PubMed: 25474690) »

    See all 2 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (HeLa cell lysate)
    Loading amount 22 µg
    Specification HeLa cell lysate
    Treatment +/- 10 Gy IR
    Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    提交于 May 13 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Blocking step 5% BSA + 1% NDS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Sample Human Cell (HeLa)
    Specification HeLa
    Permeabilization Yes - 0.1% Triton X-100
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Jul 09 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (Hela, HT29)
    Loading amount 30 µg
    Specification Hela, HT29
    Treatment see legend
    Gel Running Conditions Reduced Denaturing (7% Tris acetate)
    Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    提交于 May 02 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.5% Triton X-100 in PBS
    Username

    Dr. Kirk McManus

    Verified customer

    提交于 Feb 19 2013

    Thank you for contacting us.

    Below you can find the tested WB protocol.

    Protocols: Western Blot

    Cell Lysis and Western Blotting Protocol


    http://www.epitomics.com/pdf/westerndata.pdf

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"