Human BMI1 knockout MCF7 cell lysate (ab256851)
Overview
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Product name
Human BMI1 knockout MCF7 cell lysate
See all Bmi1 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
MCF7 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 1 bp insertion in exon2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.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.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab263505 - Human BMI1 knockout MCF7 cell lysate 1 x 100µg ab263912 - Human wild-type MCF7 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma
Target
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Function
Component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility. In the PRC1 complex, it is required to stimulate the E3 ubiquitin-protein ligase activity of RNF2/RING2. -
Sequence similarities
Contains 1 RING-type zinc finger. -
Post-translational
modificationsMonoubiquitinated (By similarity). May be polyubiquitinated; which does not lead to proteasomal degradation. -
Cellular localization
Nucleus. Cytoplasm. - Information by UniProt
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Alternative names
- B lymphoma Mo MLV insertion region (mouse)
- B lymphoma Mo MLV insertion region 1 homolog
- Bmi 1
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab256851 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 36 kDa. |
Images
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Lane 1: Wild-type MCF7 cell lysate (20µg)
Lane 2: BMI1 knockout MCF7 cell lysate (20µg)
Lane 3: A431 cell lysate (20µg)
Lane 4: HEK-293 cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - ab269678 observed at 36 kDa. Red - loading control ab52866 observed at 50 kDa.
ab269678 Mouse monoclonal [BMI1/2823] to Bmi1 was shown to specifically 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 and Bmi1 knockout samples 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) were incubated overnight at 4°C at 2 dilution and 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. -
Lane 1: Wild-type MCF7 cell lysate (20µg)
Lane 2: BMI1 knockout MCF7 cell lysate (20µg)
Lane 3: A431 cell lysate (20µg)
Lanes 1- 3: Merged signal (red and green). Green - ab126783 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab126783 Rabbit monoclonal [EPR3745(2)] to Bmi1 was shown to specifically 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 and Bmi1 knockout samples 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) were incubated overnight at 4°C at 1 in 10000 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. -
Allele-1: 1 bp deletion in exon2
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Allele-2: 1 bp insertion in exon2
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab256851 has not yet been referenced specifically in any publications.