Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade - BSA and Azide free

16 product images

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

  Bmi1 was immunoprecipitated from 0.35mg of K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/1000 dilution. VeriBlot for IP detection reagent (HRP) (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: Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 IP in K562 lysate.

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade 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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

  All lanes: Immunoprecipitation - Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253)

  Predicted band size: 36 kDa

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

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (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 (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 Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade 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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

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

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/60 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

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

  Immunohistochemical analysis of paraffin-embedded human prostatic hyperplasia tissue labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (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 (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 Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade 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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

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  ChIP - Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade - 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 Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade 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).

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

  Primers and probes are located in the first kb of the transcribed region.

• 

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

  Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (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 (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 Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade 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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

• 

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

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cell line labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/60 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

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

  Immunofluorescent analysis of 4% paraformaldehyde, 0.1% tritonX-100 fixed, NIH/3T3 (mouse embryonic fibroblast) cells labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/50 dilution, followed by AlexaFluor®488 Goat anti-Rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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, Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 was used as a counterstain.
  

  Control: Secondary antibody only

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

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade - 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 Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/50 dilution, followed by a AlexaFluor^®488 Goat anti-Rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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, Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

• 

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

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cell line labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/60 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

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

  Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8 µg of Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 [EPR22604-160]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

• 

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

  Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8 µg of Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 [EPR22604-160]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

• 

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

  Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8 µg of Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 [EPR22604-160]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

• 

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

  Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 NCCIT cells and 8 µg of Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 [EPR22604-160]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

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  ChIC/CUT&RUN sequencing - Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade - BSA and Azide free (ab254475)

  ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 NCCIT (Human pluripotent embryonic carcinoma cell line) cells and 5µg of Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 [EPR22604-160]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
  
  Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.
  
  The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
  
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).

• 

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

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Bmi1 with Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253 at 1/60 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (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 (Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade ab254253).