Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22604-160] to Bmi1 - BSA and Azide free
  • Suitable for: IHC-P, ChIP, Flow Cyt, ICC/IF, IP, WB
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free
    See all Bmi1 primary antibodies

  • Description

    Rabbit monoclonal [EPR22604-160] to Bmi1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: K562, HeLa, MOLT-4, A549 and NIH/3T3 whole cell lysates IHC-P: Human gastric cancer, prostatic hyperplasia; Mouse colon ICC/IF: HeLa and NIH/3T3 cell lines Flow Cyt: HeLa and NIH/3T3 cell lines IP: K562 cell lysate. ChIP: NCCIT cells.

  • General notes

    ab254475 is the carrier-free version of ab254253. 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

    ab254475 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • ChIP - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    ChIP - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Chromatin was prepared from NCCIT cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min.

    The ChIP was performed with 25 µg of chromatin, 5 µg of ab254253 (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Bmi1 with ab254253 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is the ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab254253 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Immunohistochemical analysis of paraffin-embedded human prostatic hyperplasia tissue labeling Bmi1 with ab254253 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human prostatic hyperplasia is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is the ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab254253 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling Bmi1 with ab254253 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human gastric cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is the ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab254253 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).

  • Immunoprecipitation - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Immunoprecipitation - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Bmi1 was immunoprecipitated from 0.35mg of K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab254253 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab254253 at 1/1000 dilution. VeriBlot for IP detection reagent (HRP) (ab131366) was used as secondary antibody at 1/5000 dilution.

    Lane 1: K562 lysate 10 μg (Input). 

    Lane 2: ab254253 IP in K562 lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab254253 in K562 whole cell lysate

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

    Exposure time: 15 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).

  • Flow Cytometry - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Flow Cytometry - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cell line labeling Bmi1 with ab254253 at 1/60 (red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).
  • Flow Cytometry - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Flow Cytometry - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Bmi1 with ab254253 at 1/60 (red) compared with a Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).
  • Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Immunofluorescent analysis of 4% paraformaldehyde, 0.1% triton X-100 fixed, NIH/3T3 (mouse embryonic fibroblast) cells labeling Bmi1 with ab254253 at 1/50 dilution, followed by AlexaFluor®488 Goat anti-Rabbit (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in NIH/3T3 cells. DAPI was used as a nuclear counterstain (blue). Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889) at 1/200 was used as a counterstain.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a AlexaFluor®488 Goat anti-Rabbit (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 (ab254253).

  • Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized, HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Bmi1 with ab254253 at 1/50 dilution, followed by a AlexaFluor®488 Goat anti-Rabbit (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in Hela cells. DAPI was used as a nuclear counterstain (blue). Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889) at 1/200 was used as a counterstain.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254253).

  • Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)
    Anti-Bmi1 antibody [EPR22604-160] - BSA and Azide free (ab254475)

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