Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

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 - ChIP Grade
  • Suitable for: ChIP, WB, ICC/IF, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade
    See all SUZ12 primary antibodies

  • Description

    Rabbit monoclonal [EPR5234(N)] to SUZ12 - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Mouse
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human SUZ12 aa 50-150 (Cysteine residue). The exact sequence is proprietary.
    Database link: Q15022

  • Positive control

    • SW480, HeLa, MCF-7 and 293T cell lysates. MCF-7 cells. Immunoprecipitation pellet from HeLa whole cell lysate (ab150035).

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: SUZ12 knockout HAP1 cell lysate (20 µg)
    Lane 3: Caco2 cell lysate (20 µg)
    Lane 4: MCF7 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab175187 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab175187 was shown to specifically react with SUZ12 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when SUZ12 knockout samples were used. Wild-type and SUZ12 knockout samples were subjected to SDS-PAGE. ab175187 and ab8245 (loading control to GAPDH) were both 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1hr at room temperature before imaging.

  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    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
  • Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    Immunofluorescence analysis of MCF-7 cells labeling SUZ12 with ab175187 at a 1/50 dilution.

  • Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    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.

  • Flow Cytometry - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Flow Cytometry - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    Flow cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SUZ12 (red) with purified ab175187 at a 1/2000 dilution (1ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    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?



    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.

  • Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Western blot - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    All lanes : Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187) at 1/1000 dilution

    Lane 1 : SW480 cell lysates
    Lane 2 : HeLa cell lysates
    Lane 3 : MCF-7 cell lysates
    Lane 4 : 293T cell lysates

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 83 kDa

  • ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    ChIP - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    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.
  • Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Immunoprecipitation - Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

    Western blot analysis on immunoprecipitation pellet from HeLa cell lysate using ab175187 at a 1/10 dilution.

  • Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)
    Anti-SUZ12 antibody [EPR5234(N)] - ChIP Grade (ab175187)

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