B2M KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1
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
B2M KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1
B2M
Knockout
CRISPR technology
Sanger Sequencing
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.
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.
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.
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Western blot: Anti-beta 2 Microglobulin antibody [EP2978Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta 2 Microglobulin antibody [EP2978Y] 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 Human B2M (beta 2 Microglobulin) knockout A-431 cell line ab261893 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout A-431 cell lysate ab261702) or B2M knockout HEK-293T cell line ab266828 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout HEK-293T 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] (Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853) at 1/5000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: B2M knockout HEK-293T cell lysate at 20 µg
Lane 3: Wild-type A431 cell lysate at 20 µg
Lane 4: B2M knockout A431 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 12 kDa
False colour image of Western blot: Anti-beta 2 Microglobulin antibody [EPR21752-214] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-beta 2 Microglobulin antibody [EPR21752-214] ab218230 was shown to bind specifically to beta 2 Microglobulin. A band was observed at 12 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in B2M knockout cell line ab266828 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout HEK-293T cell lysate ab256845). To generate this image wild-type and B2M knockout HEK-293T cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (Anti-beta 2 Microglobulin antibody [EPR21752-214] ab218230) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: B2M knockout HEK-293T cell lysate at 20 µg
Lane 3: Wild-type A431 cell lysate at 20 µg
Lane 4: B2M knockout A431 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 12 kDa
Representative images of B2M knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sequencing chromatogram displaying sequence edit in exon 1.
Homozygous: 1 bp insertion in exon 1
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