Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5234(N)] to SUZ12 - BSA and Azide free
  • Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF, ChIP
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free
    See all SUZ12 primary antibodies

  • Description

    Rabbit monoclonal [EPR5234(N)] to SUZ12 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF, ChIPmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa and HAP1 cell lysate. IP: HeLa cell lysate. ChIP: HeLa and F9 cell lysate.

  • General notes

    ab249844 is the carrier-free version of ab175187.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
    All lanes : Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SUZ12 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab175187).

      Lanes 1- 2: Merged signal (red and green). Green - ab175187 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab175187 was shown to react with SUZ12 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264983 (knockout cell lysate ab257721) lane below 100kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and SUZ12 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab175187 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
    Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)

    ab175187 (purified) at 1/20 dilution (16 µg/mL) immunoprecipitating SUZ12 in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg.
    Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
    Lane 2 (+): ab175187 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab175187 in HeLa whole cell lysate
    For western blotting, ab175187 at 1/500 dilution (0.636 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.
    Blocking and diluting buffer: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175187)

  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
    ChIP - Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)

    Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab175187 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175187).

  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
    ChIP - Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
    Chromatin was prepared from F9 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab175187 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci).
    Primers and probes 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 (ab175187).
  • Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)
    Anti-SUZ12 antibody [EPR5234(N)] - BSA and Azide free (ab249844)

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