Anti-Hu CD20 Pacific Blue™

About this product

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.

Specifications

Catalog number

PB-638-T100

Size / count

100 Tests

Dry ice

No

Hazardous

No

Discontinued

No

Certification

RUO

Reactivity

Human, Non-human primates

Immunogen

Human tonsillar B cells

Target Antigen

CD20

Clone

2H7

Applications

FC (QC tested)

Format

Pacific Blue

Buffer

Stabilizing phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide

References & Citations

*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., URL: http://www.ncbi.nlm.nih.gov/pubmed/12835728 , *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., URL: http://www.ncbi.nlm.nih.gov/pubmed/19351771 , *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., URL: http://www.ncbi.nlm.nih.gov/pubmed/18354204 , *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., URL: http://www.ncbi.nlm.nih.gov/pubmed/9417086 , *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., URL: http://www.ncbi.nlm.nih.gov/pubmed/7545683 , *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., URL: https://www.ncbi.nlm.nih.gov/pubmed/12176898, *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., URL: https://www.ncbi.nlm.nih.gov/pubmed/11964291, *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., URL: https://www.ncbi.nlm.nih.gov/pubmed/14500384, *Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995) ., URL: , *Leukocyte Typing IV., Knapp W. et al. (Eds.), Oxford University Press (1989) ., URL: , *Leukocyte Typing III., McMichael M.J. et al. (Eds.), Oxford University Press (1987) ; p.611., URL: , *Leukocyte Typing Human B Cell and B cell blast-associated Surface Molecules defined with Monoclonal Antibodies, Clerk et al., 1984, pg. 339-346., URL:

Other References

*Li H, Ayer LM, Polyak MJ, Mutch CM, Petrie RJ, Gauthier L, Shariat N, Hendzel MJ, Shaw AR, Patel KD, Deans JP. The CD20 calcium channel is localized to microvilli and constitutively associated with membrane rafts: antibody binding increases the affinity of the association through an epitope-dependent cross-linking-independent mechanism. J Biol Chem. 2004 May 7;279 (19) :19893-901., URL: https://pubmed.ncbi.nlm.nih.gov/17497959/, *Glennie MJ, French RR, Cragg MS, Taylor RP: Mechanisms of killing by anti-CD20 monoclonal antibodies. Mol Immunol. 2007 Sep;44 (16) :3823-37., URL: https://pubmed.ncbi.nlm.nih.gov/17768100/, *Cragg MS, Walshe CA, Ivanov AO, Glennie MJ: The biology of CD20 and its potential as a target for mAb therapy. Curr Dir Autoimmun. 2005;8:140-74., URL: https://pubmed.ncbi.nlm.nih.gov/15564720/, *Petrie RJ, Deans JP: Colocalization of the B cell receptor and CD20 followed by activation-dependent dissociation in distinct lipid rafts. J Immunol. 2002 Sep 15;169 (6) :2886-91., URL: https://pubmed.ncbi.nlm.nih.gov/12218101/, *Hultin LE, Hausner MA, Hultin PM, Giorgi JV: CD20 (pan-B cell) antigen is expressed at a low level on a subpopulation of human T lymphocytes. Cytometry. 1993;14 (2) :196-204., URL: https://www.ncbi.nlm.nih.gov/pubmed/7679964?dopt=A

Storage Conditions

Store at 2-8°C. Protect from prolonged exposure to light. Do not freeze.

Specificity

The mouse monoclonal antibody 2H7 recognizes an extracellular epitope on 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.

Applications Notes

6

Isotype

Mouse IgG2b kappa