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Immunofluorescence (IF) (Immunofluorescence staining of U937 cells using anti-CD20 antibody (MBS488637) 18B12. Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes immobilized on Shi-fix cover-slips and stained with the chimeric rabbit IgG version of 18B12 () at 10ug/ml followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom , DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (MBS488041) followed by Alexa Fluor 488 secondary antibody.)

Rabbit anti-Mouse CD20 Monoclonal Antibody | anti-CD20 antibody

Anti-CD20 [18B12]

Gene Names
Ms4a1; Cd20; Ly-44; Ms4a2; AA960661
Reactivity
Mouse
Applications
Flow Cytometry, Functional Assay, Immunofluorescence, Western Blot, Functional Assay
Purity
Purified antibody. Protein A affinity purified
Synonyms
CD20; Monoclonal Antibody; Anti-CD20 [18B12]; Ly-44; B-cell differentiation antigen Ly-44; B-lymphocyte antigen CD20; CD20 antigen; Lymphocyte antigen 44; Membrane-spanning 4-domains subfamily A member 1; membrane-spanning 4-domains; subfamily A; member 2; anti-CD20 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Mouse
Clonality
Monoclonal
Isotype
Rabbit IgG, kappa
Clone Number
18B12
Specificity
This antibody is specific for murine CD20, which is a cell surface 33-37 (depending on the degree of phosphorylation) kDa non-glycosylated surface phosphoprotein expressed on mature and most malignant B cells, but not stem cells or plasma cells. Low number of the CD20 has been also detected on a subpopulation of T lymphocytes and it can be expressed on follicular dendritic cells. Its expression on B cells is synchronous with the expression of surface IgM. CD20 regulates transmembrane calcium conductance (probably functioning as a component of store-operated calcium channel), cell cycle progression and B-cell proliferation. CD20 serves as a useful target for antibody-mediated therapeutic depletion of B cells, as it is expressed at high levels on most B-cell malignancies, but does not become internalized or shed from the plasma membrane following mAb treatment.
Purity/Purification
Purified antibody. Protein A affinity purified
Form/Format
PBS with 0.02% Proclin 300.
Concentration
1mg/ml (varies by lot)
Applicable Applications for anti-CD20 antibody
Flow Cytometry (FC/FACS), Immunofluorescence (IF), Western Blot (WB), depleting, Functional Assay (FA)
Application Notes
In the original publication, this antibody was used in FACS and in vivo functional assays to examine how B cell depletion affects autoimmunity (Ahuja et al., 2007). In other studies, this antibody has been used mainly in FACS, immunofluorescence, and various in vitro and in vivo functional experiments. More recent works employing this antibody have shown, for instance, that enhanced phagocytosis of circulating B cells by Kupffer cells represents an important in vivo mechanism for improved activity of glycoengineered anti-CD20 mAbs (Grandjean et al., 2016), that B lymphocytes limit senescence?driven fibrosis resolution and favor hepatocarcinogenesis in mouse liver injury (Faggioli et al., 2017), and that B cells inhibit the antitumor immunity against an established murine fibrosarcoma (Maglioco et al., 2017).
Chimeric Use Statement
This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques.
PMID
17709552
Preparation and Storage
Store at 4 degree C for up to 3 months. For longer storage, aliquot and store at -20 degree C.

Immunofluorescence (IF)

(Immunofluorescence staining of U937 cells using anti-CD20 antibody (MBS488637) 18B12. Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes immobilized on Shi-fix cover-slips and stained with the chimeric rabbit IgG version of 18B12 () at 10ug/ml followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom , DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (MBS488041) followed by Alexa Fluor 488 secondary antibody.)

Immunofluorescence (IF) (Immunofluorescence staining of U937 cells using anti-CD20 antibody (MBS488637) 18B12. Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes immobilized on Shi-fix cover-slips and stained with the chimeric rabbit IgG version of 18B12 () at 10ug/ml followed by Alexa Fluor 488 secondary antibody (2ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom , DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (MBS488041) followed by Alexa Fluor 488 secondary antibody.)

Flow Cytometry (FC/FACS)

(Flow-cytometry using the Anti-CD20 antibody 18B12 (MBS488637). Splenocytes were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the rabbit IgG version of 18B12 (, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.)

Flow Cytometry (FC/FACS) (Flow-cytometry using the Anti-CD20 antibody 18B12 (MBS488637). Splenocytes were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the rabbit IgG version of 18B12 (, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.)
Related Product Information for anti-CD20 antibody
Recombinant monoclonal antibody to CD20. Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 18B12.
This antibody was generated by immunizing CD20-/- mice with CD20-transfected cell lines (300.18 and 70Z3) and CD20-peptide-keyhole limpet hemocyanin conjugate.
Product Categories/Family for anti-CD20 antibody
References
Ahuja et al. Depletion of B cells in murine lupus: efficacy and resistance. J Immunol. 2007 Sep 1;179(5):3351-61
Note on publication: Describe the original generation of this antibody and its in vivo application to better understand how B cell depletion affects autoimmunity.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
31,958 Da
NCBI Official Full Name
B-lymphocyte antigen CD20
NCBI Official Synonym Full Names
membrane-spanning 4-domains, subfamily A, member 1
NCBI Official Symbol
Ms4a1
NCBI Official Synonym Symbols
Cd20; Ly-44; Ms4a2; AA960661
NCBI Protein Information
B-lymphocyte antigen CD20
UniProt Protein Name
B-lymphocyte antigen CD20
Protein Family
UniProt Gene Name
Ms4a1
UniProt Synonym Gene Names
Cd20; Ly-44; Ms4a2

Uniprot Description

MS4A1: This protein may be involved in the regulation of B-cell activation and proliferation. Defects in MS4A1 are the cause of immunodeficiency common variable type 5 (CVID5); also called antibody deficiency due to CD20 defect. CVID5 is a primary immunodeficiency characterized by antibody deficiency, hypogammaglobulinemia, recurrent bacterial infections and an inability to mount an antibody response to antigen. The defect results from a failure of B-cell differentiation and impaired secretion of immunoglobulins; the numbers of circulating B-cells is usually in the normal range, but can be low. Belongs to the MS4A family.

Protein type: Cell surface; Membrane protein, integral; Membrane protein, multi-pass

Chromosomal Location of Human Ortholog: 19 A|19 8.19 cM

Cellular Component: external side of plasma membrane; extracellular exosome; extracellular space; integral component of membrane; integral component of plasma membrane; membrane; plasma membrane

Molecular Function: epidermal growth factor receptor binding; protein binding

Biological Process: B cell activation

Research Articles on CD20

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Product Notes

The CD20 ms4a1 (Catalog #AAA488637) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-CD20 [18B12] reacts with Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's CD20 can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS), Immunofluorescence (IF), Western Blot (WB), depleting, Functional Assay (FA). In the original publication, this antibody was used in FACS and in vivo functional assays to examine how B cell depletion affects autoimmunity (Ahuja et al., 2007). In other studies, this antibody has been used mainly in FACS, immunofluorescence, and various in vitro and in vivo functional experiments. More recent works employing this antibody have shown, for instance, that enhanced phagocytosis of circulating B cells by Kupffer cells represents an important in vivo mechanism for improved activity of glycoengineered anti-CD20 mAbs (Grandjean et al., 2016), that B lymphocytes limit senescence?driven fibrosis resolution and favor hepatocarcinogenesis in mouse liver injury (Faggioli et al., 2017), and that B cells inhibit the antitumor immunity against an established murine fibrosarcoma (Maglioco et al., 2017). Researchers should empirically determine the suitability of the CD20 ms4a1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CD20, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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