B2M KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
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Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
Alternative names
B2M, B2MG_HUMAN, Beta 2 microglobin, Beta 2 microglobulin, Beta 2 microglobulin precursor, Beta chain of MHC class I molecules, Beta chain of mhc class 1 proteins, Beta-2-microglobulin form pI 5.3, CDABP0092, Hdcma22p, IMD43
Recommended products
B2M KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- B2M
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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
- McCoY5a + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HCT116 cell line (ab288559). 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 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Beta-2-Microglobulin (B2M) is a component of the class I major histocompatibility complex (MHC I) and plays an important role in presenting peptides to the immune system. B2M weighs approximately 11.8 kDa and is found abundantly in all nucleated cells. It has alternate names such as B2 microglobulin or beta-2-microglobulin. This protein is present in the cell membrane as a part of the MHC I which is important for immune surveillance. Additionally B2M is detectable in various biological fluids including serum and its levels can reflect physiological and pathological states.
- 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.
- Associated diseases and disorders
Beta-2-microglobulin is associated with conditions such as beta-2-microglobulin amyloidosis and certain lymphoproliferative disorders. Elevated levels of B2M in serum serve as a marker for diseases like multiple myeloma where the protein level correlates with disease severity. B2M-related amyloidosis frequently occurs in patients undergoing long-term dialysis where amyloid deposits accumulate in tissues. Linking B2M to immune system dysfunction studies have shown interactions with other proteins including components of the immune system like HLA-A and HLA-B highlighting B2M's relevance in diagnosing and understanding these conditions.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
Next Generation Sequencing - Human B2M knockout HCT116 cell line (ab286381)
4 bp deletion after Thr23 of the WT protein (allele 1) and 4 bp insertion after Lys25 of the WT protein (allele 2).
Sanger Sequencing - Human B2M knockout HCT116 cell line (ab286381)
5 bp deletion after Leu15 of the WT protein
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