Bmi1 Mouse Monoclonal Antibody [B3-G5]
cat.: EM20602
Product Type: Mouse monoclonal IgG2a, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: B3-G5
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 37 kDa
Isotype: IgG2a
Immunogen: Recombinant protein within human Bmi1 full sequence.
Positive control: Hela, HepG2, Jurkat, 293T, MCF-7, PC12, K562, NIH/3T3, LOVO, human tonsil tissue, human colon cancer tissue, human breast cancer tissue, mouse kidney tissue, human kidney tissue, human brain tissue.
Subcellular location: Nucleus, Cytoplasm
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC

1:1,000
1:200
1:50-1:100
1:100-1:200
Uniprot #: SwissProt: P35226 Human | P25916 Mouse
Entrez Gene: 307151 Rat
Alternative names: B lymphoma Mo MLV insertion region (mouse) B lymphoma Mo MLV insertion region 1 homolog Bmi 1 BMI1 BMI1 polycomb ring finger oncogene BMI1_HUMAN Flvi 2/bmi 1 FLVI2/BMI1 MGC12685 Murine leukemia viral (bmi 1) oncogene homolog Oncogene BMI 1 PCGF 4 PCGF4 Polycomb complex protein BMI 1 Polycomb complex protein BMI-1 Polycomb group protein Bmi1 Polycomb group ring finger 4 Polycomb group RING finger protein 4 RING finger protein 51 RNF 51 RNF51
Images
EM20602_1.jpg Fig1: Western blot analysis of Bmi1 on different lysates using anti-Bmi1 antibody at 1/1,000 dilution.
Positive control:
Lane 1: 293T
Lane 2: Jurkat
Lane 3: Hela
Lane 4: MCF-7
Lane 5: HepG2
Lane 6: NIH/3T3
Lane 7: PC12
Lane 8: Mouse kidney
Lane 9: Human kidney
Lane 10: K562
Lane 11: Human brain
EM20602_2.jpg Fig2: Immunocytochemistry analysis of LOVO cells labeling Bmi1 with Mouse anti-Bmi1 antibody (EM20602) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Bmi1 antibody (EM20602) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
EM20602_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling Bmi1 with Mouse anti-Bmi1 antibody (EM20602) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Bmi1 antibody (EM20602) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
EM20602_4.jpg Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling Bmi1 with Mouse anti-Bmi1 antibody (EM20602) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Bmi1 antibody (EM20602) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.
EM20602_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Bmi1 antibody. Counter stained with hematoxylin.
EM20602_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-Bmi1 antibody. Counter stained with hematoxylin.
EM20602_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using anti-BMI1 antibody. Counter stained with hematoxylin.
EM20602_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-Bmi1 antibody (EM20602) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM20602) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
EM20602_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-Bmi1 antibody (EM20602) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM20602) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
EM20602_10.jpg Fig10: Flow cytometric analysis of Hela cells with BMI1 antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Goat anti mouse IgG (FITC) was used as the secondary antibody.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.