B2M KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Western blot
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 Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- B2M
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Western blot
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
- Culture medium
- F-12K + 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
Recommended control: Human wild-type A549 cell line (ab288558). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: F-12K + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x103-1x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 6x104 cells/cm2 is recommended.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
- Do not allow the cell density to exceed 7x104 cells/cm2.
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.
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2 product images
Western blot - Human B2M knockout A549 cell line (ab286573)
Western blot: Anti-B2M antibody [EP2978Y] (Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853) 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 magenta. In Western blot, Anti-beta 2 Microglobulin antibody [EP2978Y] ab75853 was shown to bind specifically to B2M. A band was observed at 13 kDa in wild-type A549 cell lysates with no signal observed at this size in B2M knockout cell line. To generate this image, wild-type and B2M knockout A549 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 °C. 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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 A549 cell lysate at 20 µg
Lane 2: B2M knockout A549 cell lysate at 20 µg
Lane 3: Wild-type HEK-293T ab255553 cell lysate at 20 µg
Lane 4: B2M knockout HEK-293T Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line ab266828 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Next Generation Sequencing - Human B2M knockout A549 cell line (ab286573)
13 bp deletion after Val48 of the WT protein (allele 1); 2 bp deletion after Pro24 of the WT protein (allele 2)
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