Human BMI1 knockout MCF7 cell line
- Advanced Validation
- What is this?
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BMI1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2.
View Alternative Names
B lymphoma Mo MLV insertion region (mouse), B lymphoma Mo MLV insertion region 1 homolog, BMI1 polycomb ring finger oncogene, BMI1_HUMAN, Flvi 2/bmi 1, MGC12685, Murine leukemia viral (bmi 1) oncogene homolog, Oncogene BMI 1, PCGF 4, Polycomb complex protein BMI-1, Polycomb group RING finger protein 4, Polycomb group protein Bmi1, Polycomb group ring finger 4, RING finger protein 51, RNF 51
- WB
Lab
Western blot - Human BMI1 knockout MCF7 cell line (AB262319)
Lanes 1- 4 : Merged signal (red and green). Green - ab269678 observed at 36 kDa. Red - Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) observed at 50 kDa.
ab269678 was shown to react with Bmi1 in wild-type MCF7 cells in western blot. Loss of signal was observed when knockout cell line ab262319 (knockout cell lysate ab256851) was used. Wild-type MCF7 and BMI1 knockout MCF7 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab269678 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) overnight at 4°C at a 2 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Bmi1 antibody [BMI1/2823] (<a href='/en-us/products/primary-antibodies/bmi1-antibody-bmi1-2823-ab269678'>ab269678</a>) at 2 µg/mL
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 1:
Western blot - Human BMI1 knockout MCF7 cell lysate (<a href='/en-us/products/cell-lysates/human-bmi1-knockout-mcf7-cell-lysate-ab256851'>ab256851</a>)
Lane 2:
BMI1 knockout MCF7 cell lysate at 20 µg
Lane 2:
Western blot - Human BMI1 knockout MCF7 cell line (ab262319)
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
HEK-293 cell lysate at 20 µg
Predicted band size: 36 kDa
Observed band size: 36 kDa
false
- WB
Lab
Western blot - Human BMI1 knockout MCF7 cell line (AB262319)
Lanes 1- 3 : Merged signal (red and green). Green - ab126783 observed at 37 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab126783 was shown to react with Bmi1 in wild-type MCF7 cells in western blot. Loss of signal was observed when knockout cell line ab262319 (knockout cell lysate ab256851) was used. Wild-type MCF7 and BMI1 knockout MCF7 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab126783 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Bmi1 antibody [EPR3745(2)] (<a href='/en-us/products/primary-antibodies/bmi1-antibody-epr37452-ab126783'>ab126783</a>) at 1/10000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
BMI1 knockout MCF7 cell lysate at 20 µg
Lane 2:
Western blot - Human BMI1 knockout MCF7 cell line (ab262319)
Lane 3:
A431 cell lysate at 20 µg
Predicted band size: 171 kDa,36 kDa,43 kDa,67 kDa
Observed band size: 175 kDa,37 kDa,67 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human BMI1 knockout MCF7 cell line (AB262319)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human BMI1 knockout MCF7 cell line (AB262319)
Allele-1 : 1 bp deletion in exon2
Reactivity data
Product details
Recommended control: Human wild-type MCF7 cell line (ab257303). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
MEM + 10% FBS + 0.01 mg/ml bovine insulin
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
The functions of Bmi1 extend into maintaining stem cell renewal and differentiation. It acts as part of the Polycomb repressive complex 1 (PRC1) partnering with other proteins to regulate gene expression by altering chromatin structure. Bmi1 protein regulates cell proliferation and inhibits senescence by repressing the INK4a/ARF locus which encodes tumor suppressor proteins like p16INK4a and p14ARF.
Pathways
Bmi1 plays significant roles in processes such as the DNA damage response and the self-renewal of hematopoietic stem cells. It is involved in the modulation of the Wnt and Notch signaling pathways. Bmi1 protein interacts closely with proteins such as CBX2 and RNF2 within these pathways affecting the transcriptional landscape that governs cell fate decisions.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB254475
Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade - BSA and Azide free
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![Flow Cytometry (Intracellular) - Anti-Bmi1 antibody [EPR22604-160] - ChIP Grade - BSA and Azide free (AB254475)](https://content.abcam.com/products/images/bmi1-antibody-epr22604-160-chip-grade-bsa-and-azide-free-ab254475--flow-cytometry-intracellular-img38573.jpg)
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Product promise
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