Human B2M (beta 2 Microglobulin) knockout A-431 cell line
- Advanced Validation
- What is this?
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(1 Publication)
- WB
Lab
Western blot - Human B2M (beta 2 Microglobulin) knockout A-431 cell line (AB261893)
Western blot : Anti-beta 2 Microglobulin antibody [EP2978Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab75853 was shown to bind specifically to beta 2 Microglobulin. A band was observed at 12 kDa in Wild-type A431 and HEK-293T cell lysates with no signal observed at this size in B2M (beta 2 Microglobulin) knockout A-431 cell line ab261893 (knockout cell lysate ab261702) or B2M knockout HEK-293T cell line ab266828 (knockout cell lysate ab256845). To generate this image, wild-type and B2M knockout A431 and HEK-293T cell lysates were analysed. Nitrocellulose membranes were blocked in fluorescent western blot (TBS-based) blocking solution in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.
All lanes:
Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (<a href='/en-us/products/primary-antibodies/beta-2-microglobulin-antibody-ep2978y-ab75853'>ab75853</a>) at 1/5000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-b2m-beta-2-microglobulin-knockout-hek-293t-cell-lysate-ab256845'>ab256845</a>) at 20 µg
Lane 3:
Wild-type A431 cell lysate at 20 µg
Lane 4:
Western blot - Human B2M (beta 2 Microglobulin) knockout A-431 cell lysate (<a href='/en-us/products/cell-lysates/human-b2m-beta-2-microglobulin-knockout-a-431-cell-lysate-ab261702'>ab261702</a>) at 20 µg
Predicted band size: 14 kDa
Observed band size: 12 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human B2M (beta 2 Microglobulin) knockout A-431 cell line (AB261893)
23 bp deletion after Met0 (allele 1), 20 bp deletion after Arg3 (allele 2), and 1 bp deletion after Leu4 (allele 3) of the WT protein
- NGS
Lab
Next Generation Sequencing - Human B2M (beta 2 Microglobulin) knockout A-431 cell line (AB261893)
X = 23 bp deletion, 1 bp deletion, 20 bp deletion
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Cell Lines & Lysates
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Advanced Validation
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- 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
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
Beta-2-microglobulin is important for the stability and transport of MHC class I molecules to the cell surface. As part of the MHC class I complex B2M assists in binding peptides allowing immune cells to identify and target pathogen-infected cells. Without B2M the MHC class I molecules are not properly expressed on the cell surface disrupting immune recognition. In laboratory settings researchers often use anti-beta-2-microglobulin antibodies to investigate its role in MHC class I function.
Pathways
Beta-2-microglobulin interacts significantly with the immune system most notably in the antigen processing and presentation pathway. It works alongside proteins such as the heavy chain of MHC class I. B2M is important in the pathway that involves the transport of antigens to the endoplasmic reticulum where they are loaded onto MHC class I molecules for inspection by cytotoxic T cells. Another related pathway is the tapasin-mediated processing of antigen peptides highlighting the indispensable role of B2M in immune response regulation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Molecular cancer therapeutics 24:331-344 PubMed39301613
2024
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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