Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Cdk8 antibody [EPR21005] (ab229192)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21005] to Cdk8
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Cdk8 antibody [EPR21005]
    See all Cdk8 primary antibodies

  • Description

    Rabbit monoclonal [EPR21005] to Cdk8

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HeLa, Daudi, HAP1, HCT 116, K562 and SW480 whole cell lysates; Human testis lysate. IP: SW480 whole cell 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
    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.

Properties

Images

  • Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)
    Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)
    All lanes : Anti-Cdk8 antibody [EPR21005] (ab229192) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : Cdk8  knockout HeLa cell lysate
    Lane 3 : HAP1 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 53 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab229192 observed at 53 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab229192 Anti-Cdk8 antibody [EPR21005] was shown to specifically react with Cdk8 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265087 (knockout cell lysate ab257885) was used. Wild-type and Cdk8 knockout samples were subjected to SDS-PAGE. ab229192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunoprecipitation - Anti-Cdk8 antibody [EPR21005] (ab229192)
    Immunoprecipitation - Anti-Cdk8 antibody [EPR21005] (ab229192)

    Cdk8 was immunoprecipitated from 0.35 mg of SW480 (human colorectal adenocarcinoma cell line) whole cell lysate with ab229192 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229192 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/10,000 dilution.

    Lane 1: SW480 whole cell lysate 10 μg (Input).
    Lane 2: ab229192 IP in SW480 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229192 in SW480 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

  • Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)
    Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)
    All lanes : Anti-Cdk8 antibody [EPR21005] (ab229192) at 1/1000 dilution

    Lane 1 : Human testis lysate
    Lane 2 : K562 (human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
    Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 58 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)
    Western blot - Anti-Cdk8 antibody [EPR21005] (ab229192)
    All lanes : Anti-Cdk8 antibody [EPR21005] (ab229192) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate at 20 µg
    Lane 2 : Cdk8 knockout HAP1 whole cell lysate at 20 µg
    Lane 3 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate at 10 µg
    Lane 4 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 48 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

    ab229192 was shown to specifically react with Cdk8 in wild-type HAP1 cells as signal was lost in Cdk8 knockout cells.

    Wild-type and Cdk8 knockout samples were subjected to SDS-PAGE. ab229192 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

  • Anti-Cdk8 antibody [EPR21005] (ab229192)
    Anti-Cdk8 antibody [EPR21005] (ab229192)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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