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AB273825

Human MS4A1 knockout Raji cell lysate

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MS4A1 KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift: 100%.

View 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

4 Images
Western blot - Human MS4A1 knockout Raji cell lysate (AB273825)
  • WB

Lab

Western blot - Human MS4A1 knockout Raji cell lysate (AB273825)

Lane 1 : Wild-type Raji cell lysate 20 μg
Lane 2 : MS4A1 knockout Raji cell lysate 20 μg
Lane 3 : A549 cell lysate 20 μg
Lane 4 : HeLa cell lysate 20 μg

False colour image of Western blot : Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) staining, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 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°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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-mouse-igg1-chimeric-ab279298'>ab279298</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-ab78237'>ab78237</a>) 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 (<a href='/en-us/products/cell-lines/human-ms4a1-cd20-knockout-raji-cell-line-ab273871'>ab273871</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 21 kDa,33 kDa

Observed band size: 33 kDa

false

Western blot - Human MS4A1 knockout Raji cell lysate (AB273825)
  • WB

Lab

Western blot - Human MS4A1 knockout Raji cell lysate (AB273825)

Lane 1 : Wild-type Raji cell lysate 20 μg
Lane 2 : MS4A1 knockout Raji cell lysate 20 μg
Lane 3 : A549 cell lysate 20 μg
Lane 4 : HeLa cell lysate 20 μg

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] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 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 °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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-ab78237'>ab78237</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-mouse-igg2a-chimeric-ab279299'>ab279299</a>) 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 (<a href='/en-us/products/cell-lines/human-ms4a1-cd20-knockout-raji-cell-line-ab273871'>ab273871</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 21 kDa,33 kDa

Observed band size: 33 kDa

false

Western blot - Human MS4A1 knockout Raji cell lysate (AB273825)
  • WB

Lab

Western blot - Human MS4A1 knockout Raji cell lysate (AB273825)

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-ab78237'>ab78237</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) (<a href='/en-us/products/primary-antibodies/cd20-antibody-ep459y-rat-igg2a-chimeric-ab279300'>ab279300</a>) 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 (<a href='/en-us/products/cell-lines/human-ms4a1-cd20-knockout-raji-cell-line-ab273871'>ab273871</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 21 kDa,33 kDa

Observed band size: 33 kDa

false

Next Generation Sequencing - Human MS4A1 knockout Raji cell lysate (AB273825)
  • NGS

Supplier Data

Next Generation Sequencing - Human MS4A1 knockout Raji cell lysate (AB273825)

Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift : 100%

Key facts

Cell type

Raji

Species or organism

Human

Tissue

Lymphatic

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 2 bp deletion; Frameshift: 100%

Disease

Lymphoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab273825-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human wild-type Raji cell lysate", "number":"AB273825-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human MS4A1 knockout Raji cell lysate", "number":"AB273825-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
MS4A1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Suspension

Gender

Male

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Product protocols

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Product promise

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