B2M KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.
Hep G2
Human
Liver
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette 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. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1.
Hep G2
Human
Liver
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and Insertion of the selection cassette in exon 1
Hepatocellular Carcinoma
B2M
Knockout
CRISPR technology
Sanger Sequencing, Western blot
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 1 US: 1
Adherent
Male
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.
MEM + 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 HepG2 cell line (Human wild-type Hep G2 cell line ab257304). 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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Lanes 1-4: Merged signal (red and green). Green - Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853 observed at 14 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853 Anti-beta 2 Microglobulin antibody [EP2978Y] was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line ab262325 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout Hep G2 cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-beta 2 Microglobulin antibody [EP2978Y] (Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853) at 1/1000 dilution
Lane 1: Wild-type HepG2 cell lysate at 20 µg
Lane 2: B2M knockout HepG2 cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDa
This data was developed using the same antibody clone in a different buffer formulation (Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853).
Lanes 1-4: Merged signal (red and green). Green - Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853 observed at 14 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853 Anti-beta 2 Microglobulin antibody [EP2978Y] was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line ab262325 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout Hep G2 cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-2: Insertion of the selection cassette in exon 1.
Allele-1: 1 bp insertion in exon1
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