Recombinant
RabMAb

Recombinant Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - BSA and Azide free (ab232318)

Overview

  • Product name

    Anti-KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY antibody [EPR18653] - BSA and Azide free
  • Description

    Rabbit monoclonal [EPR18653] to KDM5C / Jarid1C / SMCX + KDM5D / Jarid1D / SMCY - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ChIP, IPmore 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: Jurkat, HeLa, HEL, MCF7, PC-12, NIH/3T3 and F9 whole cell lysates.
  • General notes

    Ab232318 is the carrier-free version of ab194288. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab232318 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab232318 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
WB Use at an assay dependent concentration. Detects a band of approximately 174 kDa (predicted molecular weight: 174 kDa).
ChIP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

Images

  • 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 at 20 µg
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
    Lane 3 : HEL (Human bone marrow erythroleukemia cell line) whole cell lysate at 20 µg
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg/ml

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194288).

References

ab232318 has not yet been referenced specifically in any publications.

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