Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19848] to Bmi1 (phospho T275) - BSA and Azide free
  • Suitable for: WB, Dot blot, ICC/IF, Flow Cyt, IP
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free
    See all Bmi1 primary antibodies

  • Description

    Rabbit monoclonal [EPR19848] to Bmi1 (phospho T275) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Dot blot, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • Positive control

    • ICC/IF: HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 (exposed to UV light).

  • General notes

    Ab228458 is the carrier-free version of ab213723. 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

    ab228458 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    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

  • Immunoprecipitation - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)
    Immunoprecipitation - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)

    Bmi1 (phospho T275) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector, followed by treatment with 800 J/m² UV, then cultured in DMEM containing 10% FBS for 30 minutes) whole cell lysate with ab213723 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213723 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T (transfected / UV treated) whole cell lysate 10 μg (Input).

    Lane 2: ab213723 IP in HEK-293T (transfected / UV treated) lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213723 in HEK-293T (transfected / UV treated) whole cell lysate.

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

    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213723).

  • Flow Cytometry - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)
    Flow Cytometry - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line transfected with DDDDK and mCherry-tagged mouse Bmi1 T275A mutant expression vector (left), or DDDDK and mCherry-tagged mouse Bmi1 WT expression vector (right) labeling Bmi1 (phospho T275) with ab213723 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (black). 

    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

    The cells were gated on the mCherry positive population.

    The plasmids were kindly provided by Dr. Wei Guo from Tsinghua University.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213723).

  • Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)
    Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-2 OS (human bone osteosarcoma epithelial cell line) cells labeling Bmi1 (phospho T275) with ab213723 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased nuclear foci staining observed in UV-treated U-2 OS cells. The cells were treated with 50 J/m² UV, then cultured in McCoy's 5a media supplemented with 10% FBS for 2 hours.

    The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)  at 1/200 dilution (red).

    The negative controls are as follows:
    -ve control 1: PBS instead of primary antibody (non-treated cells), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
    -ve control 2: PBS instead of primary antibody (UV treated cells), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213723).

  • Dot Blot - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)
    Dot Blot - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)

    Dot blot analysis of Bmi1 (phospho T275) labeled with ab213723 at 1/1,000 dilution.

    Lane 1: Bmi1 (phospho T275) peptide.

    Lane 2: Bmi1 non-phospho peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100,000 dilution was used as secondary antibody.

    Blocking and dilution buffer: 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 (ab213723).

  • Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)
    Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T cells (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector labeling Bmi1 (phospho T275) with ab213723 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 (WT) expression vector. No staining was observed in HEK-293T cells transfected with DDDDK and mCherry-tagged mouse Bmi1 T275A mutant expression vector.

    The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (Alexa Fluor® 647) (ab195884) at 1/200 dilution (white).

    The negative controls are as follows:
    -ve control 1: PBS instead of primary antibody (HEK-293T transfected with wild-type BMi1), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
    -ve control 2: PBS instead of primary antibody (HEK-293T transfected with Bmi1 T275A construct), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    Plasmids were kindly provided by Dr. Wei Guo from Tsinghua University.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213723).

  • Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)
    Anti-Bmi1 (phospho T275) antibody [EPR19848] - BSA and Azide free (ab228458)

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