Recombinant
RabMAb

Recombinant Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)

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Overview

  • Product name

    Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade

  • Description

    Rabbit monoclonal [EPR18653] to KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ChIP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human KDM5D/ Jarid1D/ SMCY aa 1-200. The exact sequence is proprietary.
    Database link: Q9BY66

  • Positive control

    • WB: His-tagged human KDM5D recombinant protein fragment, aa1-200; His-tagged human KDM5C recombinant protein fragment, aa1-200; Jurkat, HeLa, HEL, MCF7, PC-12, NIH/3T3 and F9 whole cell lysates. IP: HeLa and F9 whole cell lysates. ChIP: Chromatin from HeLa cells.

  • 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

Applications

Our Abpromise guarantee covers the use of ab194288 in the following tested applications.

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

Application Abreviews Notes
IP 1/50.
ChIP Use 5 µg for 25 µg of chromatin.
WB 1/1000. Detects a band of approximately 174 kDa (predicted molecular weight: 174 kDa).

Target

Images

  • ChIP - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    ChIP - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)

    Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25μg of chromatin, 5μg of ab194288 (blue), and 20μl of Anti-rabbit IgG sepharose beads. 5µg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach). Primers are located in the first kb of the transcribed region.

  • Western blot - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    Western blot - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    All lanes : Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288) at 1/5000 dilution

    Lane 1 : His-tagged human KDM5D recombinant protein fragment, aa1-200
    Lane 2 : His-tagged human KDM5C recombinant protein fragment, aa1-200

    Lysates/proteins at 0.01 µg per lane.

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

    Predicted band size: 174 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The predicted molecular weight of KDM5D recombinant protein fragment is 24.5 kDa. The predicted molecular weight of KDM5C recombinant protein fragment is 25 kDa

  • Western blot - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    Western blot - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    All lanes : Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288) at 1/1000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : HEL (Human bone marrow erythroleukemia cell line) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 174 kDa
    Observed band size: 174 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    WB detection of KDM5C + KDM5D in tissue lysates may need optimization. In our hands this antibody could not detect KDM5C + KDM5D in human tissue lysates in WB.

  • Western blot - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    Western blot - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    All lanes : Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288) at 1/1000 dilution

    Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 3 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 174 kDa
    Observed band size: 174 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 3 minutes; Lane 3: 15 seconds.

    WB detection of KDM5C + KDM5D in tissue lysates may need optimization. In our hands this antibody could not detect KDM5C + KDM5D in mouse and rat tissue lysates in WB.

  • Immunoprecipitation - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    Immunoprecipitation - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)

    KDM5D + KDM5C was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab194288 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab194288 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10 μg (Input).

    Lane 2: ab194288 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194288 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunoprecipitation - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)
    Immunoprecipitation - Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - ChIP Grade (ab194288)

    KDM5D + KDM5C was immunoprecipitated from 0.35 mg of F9 (Mouse embryonic testicular cancer cell line) whole cell lysate with ab194288 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab194288 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: F9 whole cell lysate 10 μg (Input).

    Lane 2: ab194288 IP in F9 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab194288 in F9 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

References

Publishing research using ab194288? Please let us know so that we can cite the reference in this datasheet.

ab194288 has not yet been referenced specifically in any publications.

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