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Mouse Monoclonal to CD20

2H7 (IgG2b)

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Background

CD20 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. It is associated with lipid rafts, but the intensity of this association depends on extracellular triggering, employing CD20 conformational change and/or BCR (B cell antigen receptor) aggregation. After the receptor ligation, BCR and CD20 colocalize and then rapidly dissociate before BCR endocytosis, whereas CD20 remains at the cell surface. 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.

Specificity

The mouse monoclonal antibody 2H7 recognizes CD20 (B1, Bp35), a 33-37 kDa non-glycosylated membrane receptor with four transmembrane domains, expressed on pre-B lymphocytes, resting and activated B cells (not plasma cells), follicular dendritic cells, and at low levels on peripheral blood T lymphocytes.
HLDA IV; WS Code B201

Regulatory Status

Immunogen

Human tonsillar B cells

Species Reactivity:

  • Human
  • Non-Human Primates

Negative Species:

Applications:

  • Flow Cytometry
    Recommended dilution:0.6 μg/ml
  • Immunoprecipitation
  • Immunohistochemistry (frozen sections)
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

Product Specific References

  • *Leukocyte Typing Human B Cell and B cell blast-associated Surface Molecules defined with Monoclonal Antibodies, Clerk et al., 1984, pg. 339-346.
  • *Leukocyte Typing III., McMichael M.J. et al. (Eds.), Oxford University Press (1987); p.611.
  • *Leukocyte Typing IV., Knapp W. et al. (Eds.), Oxford University Press (1989).
  • *Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995).
  • *Chan HT, Hughes D, French RR, Tutt AL, Walshe CA, Teeling JL, Glennie MJ, Cragg MS: CD20-induced lymphoma cell death is independent of both caspases and its redistribution into triton X-100 insoluble membrane rafts. Cancer Res. 2003 Sep 1;63(17):5480-9. [Abstract] [Full Text]
  • *Polyak MJ, Deans JP: Alanine-170 and proline-172 are critical determinants for extracellular CD20 epitopes; heterogeneity in the fine specificity of CD20 monoclonal antibodies is defined by additional requirements imposed by both amino acid sequence and quaternary structure. Blood. 2002 May 1;99(9):3256-62. [Abstract] [Full Text]
  • *Rose AL, Smith BE, Maloney DG: Glucocorticoids and rituximab in vitro: synergistic direct antiproliferative and apoptotic effects. Blood. 2002 Sep 1;100(5):1765-73. [Abstract] [Full Text]
  • *Deans JP, Kalt L, Ledbetter JA, Schieven GL, Bolen JB, Johnson P: Association of 75/80-kDa phosphoproteins and the tyrosine kinases Lyn, Fyn, and Lck with the B cell molecule CD20. Evidence against involvement of the cytoplasmic regions of CD20. J Biol Chem. 1995 Sep 22;270(38):22632-8. [Abstract] [Full Text]
  • *Deans JP, Robbins SM, Polyak MJ, Savage JA: Rapid redistribution of CD20 to a low density detergent-insoluble membrane compartment. J Biol Chem. 1998 Jan 2;273(1):344-8. [Abstract] [Full Text]
  • *Diehl SA, Schmidlin H, Nagasawa M, van Haren SD, Kwakkenbos MJ, Yasuda E, Beaumont T, Scheeren FA, Spits H: STAT3-mediated up-regulation of BLIMP1 Is coordinated with BCL6 down-regulation to control human plasma cell differentiation. J Immunol. 2008 Apr 1;180(7):4805-15. [Abstract] [Full Text]
  • *Hayden-Ledbetter MS, Cerveny CG, Espling E, Brady WA, Grosmaire LS, Tan P, Bader R, Slater S, Nilsson CA, Barone DS, Simon A, Bradley C, Thompson PA, Wahl AF, Ledbetter JA: CD20-directed small modular immunopharmaceutical, TRU-015, depletes normal and malignant B cells. Clin Cancer Res. 2009 Apr 15;15(8):2739-46. [Abstract] [Full Text]
  • *Polyak MJ, Ayer LM, Szczepek AJ, Deans JP: A cholesterol-dependent CD20 epitope detected by the FMC7 antibody. Leukemia. 2003 Jul;17(7):1384-9. [Abstract] [Full Text]
  • *And many other.
  • For research use only. Not for drug, diagnostic or other use.

    Example Data

    2H7 PE

    Fig. 1. Surface staining of human peripheral blood with anti-CD20 (2H7) PE.


    2H7 FITC

    Fig. 2. Surface staining of human peripheral blood with anti-CD20 (2H7) FITC.


    2H7 PE-Cy7 FC

    Fig. 3. Surface staining of human peripheral blood (lymphocyte gate) with anti-CD20 (2H7) PE-CyTM7.


    2H7 PE-Cy5 FC

    Fig. 4. Surface staining of human peripheral blood (lymphocyte gate) with anti-CD20 (2H7) PE-CyTM5.



    2H7 AF700

    Fig. 5. Surface staining of human peripheral blood with anti-CD20 (2H7) Alexa Fluor® 700.


    2H7 APC-Cy7

    Fig. 6. Surface staining of human peripheral blood with anti-CD20 (2H7) APC-CyTM7.


    2H7 PB

    Fig. 7. Surface staining of human peripheral blood with anti-CD20 (2H7) Pacific BlueTM.


    2H7 PerCP-Cy5-5

    Fig. 8. Surface staining of human peripheral blood with anti-CD20 (2H7) PerCP-CyTM5.5.


    2H7 PE-DyLight 594

    Fig. 9. Surface staining of human peripheral blood with anti-CD20 (2H7) PE-DyLigt® 594. 


    2h7 PerCP

    Fig. 10. Surface staining of human peripheral blood with anti-CD20 (2H7) PerCP.  


    2H7 APC

    Fig. 11. Surface staining of human peripheral blood with anti-CD20 (2H7) APC.  

    Unless indicated otherwise, all products are For Research Use Only and not for diagnostic or therapeutic use. Not for resale or transfer either as a stand-alone product or as a component of another product without written consent of EXBIO. EXBIO will not be held responsible for patent infringement or other violations that may occur with the use of our products. All orders are accepted subject to EXBIO´s term and conditions which are available at www.exbio.cz.
    Alexa Fluor®, Pacific Blue™ and Pacific Orange™ are registred trademarks of Molecular Probes Inc., Eugene, OR, USA. (licence label)
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    Cy™ and CyDye™ are registered trademarks of GE Healthcare.
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