MS4A1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift: 100%.
Images
Key facts
- Cell type
- Raji
- Species or organism
- Human
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift: 100%
Alternative names
APY, ATOPY, B-lymphocyte antigen CD20, B-lymphocyte cell-surface antigen B1, B-lymphocyte surface antigen B1, B1, Bp 35, CD20 antigen, CD20 receptor, CD20_HUMAN, CVID 5, FCER1B, Fc Fragment of IgE high affinity I receptor for beta polypeptide, Fc epsilon receptor I beta chain, High affinity immunoglobulin epsilon receptor subunit beta, IGEL, IGER, IGHER, IgE Fc receptor subunit beta, Leu-16, Leukocyte surface antigen Leu-16, Ly44, MGC3969, MS4A1, Membrane spanning 4 domains subfamily A member 2, Membrane-spanning 4-domains subfamily A member 1, S7, membrane-spanning 4-domains A1
Recommended products
MS4A1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift: 100%.
Key facts
- Cell type
- Raji
- Species or organism
- Human
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift: 100%
- Disease
- Lymphoma
- Concentration
Properties
- Gene name
- MS4A1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Suspension
- 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 for bath 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 4x105 - 5x105 cells/mL(for initial passages it is recomended to culture the cells in the higher range of recomended seeding density). 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 4x105 cells/mL is recommended.
- A maximum of 3x106 viable cells/mL is obtainable.
- Culture medium
- RPMI + 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
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
CD20 also known as MS4A1 or L26 is a membrane-spanning 4-domains subfamily A member 1 protein. It is a non-glycosylated phosphoprotein with a molecular mass of approximately 33-37 kDa. Expressed extensively on the surface of B-cells CD20 plays an important role in B-cell activation and regulation. While CD20 is absent on early pro-B cells and plasma cells its presence increases as B-cells mature. Anti-CD20 antibodies such as 2H7 are commonly used in research and treatment to deplete B-cells due to the target's consistent expression in most stages of B-cell development.
- Biological function summary
CD20 contributes significantly to calcium ion transport which is essential for the activation process of B-cells. Although CD20 does not belong to a larger protein complex its role centers around forming a calcium channel that allows a sustained influx of calcium into the B-cells. This influx is important for the signaling pathways that govern B-cell activation proliferation and differentiation impacting the immune response efficacy.
- Pathways
CD20 is closely involved in the B-cell receptor (BCR) signaling pathway and the Fc gamma R-mediated phagocytosis pathway. These pathways play vital roles in adaptive immunity and immune response modulation. CD20's interaction with proteins like PI3K during BCR signaling enhances receptor-mediated cellular signals subsequently influencing downstream effectors involved in cell growth and survival of B-cells.
- Associated diseases and disorders
CD20 is strongly linked to certain blood cancers and autoimmune diseases including non-Hodgkin's lymphoma and rheumatoid arthritis. These conditions often exploit aberrant B-cell activity or count. CD20's expression on malignant B-cells in non-Hodgkin's lymphoma makes it an effective target for monoclonal antibody therapies such as rituximab an anti-CD20 antibody. Additionally CD20-targeted therapies can help deplete pathogenic B-cells in rheumatoid arthritis highlighting the protein's relevance in disease treatment.
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5 product images
Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
False colour image of Western blot: Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) staining at 1/1000 dilution shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) ab279300 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line ab273871 (knockout cell lysate Human MRPS28 knockout HEK-293T cell lysate ab263259). To generate this image wild-type and MS4A1 knockout Raji cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1% Tween® 20 (TBS-T) 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-Rat IgG H&L (IRDye® 800CW) preabsorbed (ab253031) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] (Anti-CD20 antibody [EP459Y] ab78237) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) (Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) ab279300) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: MS4A1 knockout Raji cell lysate at 20 µg
Lane 2: Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
Lane 3: A549 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 21 kDa, 33 kDa
Observed band size: 33 kDa
Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
False colour image of Western blot: Anti-CD20 antibody [EP459Y] – Mouse IgG2a (Chimeric) staining at 1/1000 dilution shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) ab279299 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line ab273871 (knockout cell lysate Human MRPS28 knockout HEK-293T cell lysate ab263259). To generate this image wild-type and MS4A1 knockout Raji cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1% Tween® 20 (TBS-T) 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] (Anti-CD20 antibody [EP459Y] ab78237) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) (Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) ab279299) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: MS4A1 knockout Raji cell lysate at 20 µg
Lane 2: Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
Lane 3: A549 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 21 kDa, 33 kDa
Observed band size: 33 kDa
Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
False colour image of Western blot: Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) staining shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) ab279298 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line ab273871 (knockout cell lysate Human MRPS28 knockout HEK-293T cell lysate ab263259). To generate this image wild-type and MS4A1 knockout Raji cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) (Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) ab279298) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] (Anti-CD20 antibody [EP459Y] ab78237) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: MS4A1 knockout Raji cell lysate at 20 µg
Lane 2: Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
Lane 3: A549 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 21 kDa, 33 kDa
Observed band size: 33 kDa
Next Generation Sequencing - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift: 100%
Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
All lanes: Western blot - Anti-CD20 antibody [SP32] (Anti-CD20 antibody [SP32] ab64088) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate
Lane 2: MS4A1 knockout Raji cell lysate
Lane 2: Western blot - Human MS4A1 (CD20) knockout Raji cell line (ab273871)
Lane 3: Ramos cell lysate
Lane 4: A549 cell lysate
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
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