EM20602

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:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
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:	K562, 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	1:2,000 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

	Fig1: Western blot analysis of Bmi1 on K-562 cell lysate with Mouse anti-Bmi1 antibody (EM20602) at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 15 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: EM20602, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃ Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature Predicted band size: 37 kDa Observed band size: 37 kDa
	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.
	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.

	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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Bmi1 antibody. Counter stained with hematoxylin.
	Fig6: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-Bmi1 antibody. Counter stained with hematoxylin.
	Fig7: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using anti-BMI1 antibody. Counter stained with hematoxylin.

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) (high pressure) 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.

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) (high pressure) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.