Anti-Chk1 (phospho S345)抗体(ab58567)

概述

  • 产品名称
    Anti-Chk1 (phospho S345)抗体
    参阅全部 Chk1 一抗
  • 描述
    兔多克隆抗体to Chk1 (phospho S345)
  • 特异性
    Specific for Chk1 (phospho S345) in ELISA, with <1% reactivity with the non-modified form of the immunogen.
  • 经测试应用
    适用于: WB, IHC-P, ELISAmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse, Rat, Chicken, Cow
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 300 - 400 of Human Chk1.

    (Peptide available as ab93794.)

  • 阳性对照
    • This antibody gave a positive signal in Hydroxyurea Treated (48hr, 2uM) Hela whole cell lysate. IHC-P: FFPE Human cervix normal.
  • 常规说明

    Please note we cannot guarantee ICC/IF, however should you use this application for this product then please use a specific protocol which can be found in the protocols section of our datasheet here.

性能

应用

Our Abpromise guarantee covers the use of ab58567 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.

靶标

图片

  • All lanes : Anti-Chk1 (phospho S345) antibody (ab58567) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate - Untreated
    Lane 2 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 2uM)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 54 kDa
    Observed band size : 54 kDa
    Additional bands at : 105 kDa,42 kDa,90 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab58567 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • IHC image of ab58567 staining Human Cervical Carcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab58567, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

文献

This product has been referenced in:
  • Li B  et al. Resveratrol sequentially induces replication and oxidative stresses to drive p53-CXCR2 mediated cellular senescence in cancer cells. Sci Rep 7:208 (2017). Read more (PubMed: 28303009) »
  • Murai J  et al. Resistance to PARP inhibitors by SLFN11 inactivation can be overcome by ATR inhibition. Oncotarget 7:76534-76550 (2016). WB ; Human . Read more (PubMed: 27708213) »

See all 3 Publications for this product

客户评价及客户问答

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 line)
Loading amount
40 µg
Specification
HeLa cell line
Treatment
mitomycin C
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Sep 25 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MRC5VA Lung Epithelial Fibroblasts)
Loading amount
15 µg
Specification
MRC5VA Lung Epithelial Fibroblasts
Treatment
30GY Ionising Radiation
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Apr 13 2011

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

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