CyFlow™ CD20 Biotin

CyFlow™ CD20 Biotin
Target Species: Human
Antigen: CD20
Field of Interest: Immunophenotyping, Signaling
Clone: LT20
Regulatory Status: RUO
Alternative Name: B1, Bp35
Application: Flow cytometry
Format/Fluorochrome: Biotin
Species of Origin: Mouse
Clonality: monoclonal
Isotype: IgG2a
Product number: CL990222

For Research Use Only

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Concentration Unit mg/mL Concentration 1 Quantity 0.1 mg Immunogen Normal human... more
CyFlow™ CD20 Biotin
Concentration Unitmg/mL
Concentration1
Quantity0.1 mg
ImmunogenNormal human lymphocytes from lymph node
SpecifityThe mouse monoclonal antibody LT20 recognizes CD20 antigen, 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.
Background InformationCD20 (B1, Bp35) 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.
UsageThe reagent is designed for indirect immunofluorescence analysis by Flow Cytometry. Suggested working usage is 6.7·µg/ml. Indicated dilution is recommended starting point for use of this product, but working concentrations should be validated by the investigator.
Storage BufferThe reagent is provided in phosphate buffered saline (PBS) solution, pH ≈7.4, containing 0.1% (w/v) sodium azide.
StorageAvoid prolonged exposure to light. Store in the dark at 2-8°C. Do not freeze.
StabilityDo not use after expiration date stamped on vial label.

Specific References

| 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. <·PMID:·7523045·> | Mason D, Simmons D, Buckley C, Schwartz-Albiez R, Hadam M, Saalmuller A, Clark E, Malavasi F, Morrissey JA (Eds): Leucocyte Typing VII. Oxford·University·Press. 2002; 1‑945. <·NLM·ID:·101177131·> | 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. <·PMID:·11964291·> | 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. <·PMID:·12218101·> | 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. <·PMID:·14500384·> | 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. <·PMID:·14976189·> | 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. <·PMID:·15564720·> | Teeling JL, Mackus WJ, Wiegman LJ, van den Brakel JH, Beers SA, French RR, van Meerten T, Ebeling S, Vink T, Slootstra JW, Parren PW, Glennie MJ, van de Winkel JG: The biological activity of human CD20 monoclonal antibodies is linked to unique epitopes on CD20. J·Immunol. 2006·Jul·1; 177(1):362‑71. <·PMID:·16785532·> | Filatov AV, Krotov GI, Zgoda VG, Volkov Y: Fluorescent immunoprecipitation analysis of cell surface proteins: a methodology compatible with mass‑spectrometry. J·Immunol·Methods. 2007·Jan·30; 319(1‑2):21‑33. <·PMID:·17098248·> | Glennie MJ, French RR, Cragg MS, Taylor RP: Mechanisms of killing by anti‑CD20 monoclonal antibodies. Mol·Immunol. 2007·Sep; 44(16):3823‑37. <·PMID:·17768100·> | Kanderova V, Kuzilkova D, Stuchly J, Vaskova M, Brdicka T, Fiser K, Hrusak O, Lund‐Johansen F, Kalina T: High‐resolution antibody array analysis of childhood acute leukemia cells. Mol·Cell·Proteomics. 2016·Apr·1; 15(4):1246‐61. <·PMID:·26785729·>

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