Human BMI1 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human BMI1 knockout HEK-293T cell line (AB266514)
Lanes 1-3 : Merged signal (red and green). Green - ab126783 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab126783 Anti-Bmi1 antibody [EPR3745(2)] was shown to specifically react with Bmi1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266514 (knockout cell lysate ab256850) was used. Wild-type and Bmi1 knockout samples were subjected to SDS-PAGE. ab126783 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 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 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
BMI1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human BMI1 knockout HEK-293T cell line (ab266514)
Lane 3:
MOLT-4 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 37 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human BMI1 knockout HEK-293T cell line (AB266514)
Homozygous : 543 bp deletion in exon2
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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-img415867.jpg)
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Cell Lines & Lysates
AB264667
Human CSNK1D (Casein Kinase 1 delta) knockout HeLa cell line
Advanced Validation
- 0
- 0
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
Zygosity
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
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com