Biolegend, 302365, Spark NIR™ 685 anti-human CD20 Antibody, 25tests

CD20 is a 33-37 kD, four transmembrane spanning protein, also known as B1 and Bp35. CD20 is expressed on pre-B-cells, resting and activated B cells (not plasma cells), some follicular dendritic cells, and at low levels on a T cell subset. CD20 is heavily phosphorylated on activated B cells and malignant B cells. Homo-oligomeric complexes of CD20 are thought to form Ca 2+ conductive ion channels in the plasma membrane of B cells. The CD20 molecule is involved in B-cell activation and is associated with various Src family kinases (Lyn, Lck, Fyn). It exists in a complex with MHC class I and II, CD53, CD81, and CD82.
25tests
Verified Reactivity: Human, Cynomolgus, Rhesus
Reported Reactivity: Baboon, Capuchin Monkey, Chimpanzee, Pigtailed Macaque, Squirrel Monkey
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Human tonsillar B cells
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation: The antibody was purified by affinity chromatography and conjugated with Spark NIR™ 685 under optimal conditions.
Concentration: Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: FC - Quality tested
Recommended Usage: Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µL per million cells in 100 µL staining volume or 5 µL per 100 µL of whole blood. It is recommended that the reagent be titrated for optimal performance for each application. * Spark NIR™ 685 has a maximum excitation of 665 nm and a maximum emission of 685 nm.
Excitation Laser: Red Laser (633 nm)
Application Notes: The epitope recognized by clone 2H7 has been mapped to the sequence YNCEPANPSEKNSPST which lies in the large extracellular loop of human CD20. Additional reported applications (for the relevant formats) include: immunoprecipitation4 and immunohistochemical staining of acetone-fixed frozen sections5.
Application References(PubMed link indicates BioLegend citation): Schlossman S, et al. 1995. Leucocyte Typing V. Oxford University Press. New York. Knapp W, et al. 1989. Leucocyte Typing IV. Oxford University Press. New York. McMichael A, et al. Eds. 1987. Leucocyte Typing III Oxford University Press. New York. Polyak MJ, et al. 2002. Blood 99:3256. (IP) Mack CL, et al. 2004. Pediatr. Res. 56:79. (IHC)
RRID: AB_2860774 (BioLegend Cat. No. 302365) AB_2860775 (BioLegend Cat. No. 302366)
Structure: Four transmembrane protein (TM4SF), heavily phosphorylated after activation, 33-37 kD
Distribution: B cell, T cell subsets
Function: B cell activation
Ligand/Receptor: Src family tyrosine kinases, MHC class I, II, CD53, CD81, CD82
Cell Type: B cells, T cells
Biology Area: Costimulatory Molecules, Immunology
Molecular Family: CD Molecules
Antigen References: 1. Hultin L, et al. 1993. Cytometry 14:196. 2. Tedder T, et al. 1994. Immunol. Today 15:450.
Gene ID: 931
UniProt: View information about CD20 on UniProt.org
Clone: 2H7
Regulatory Status: RUO
Workshop: IV B201
Other Names: B1, Bp35
Isotype: Mouse IgG2b, κ
Q: What is the difference between two anti human CD20 clones 2H7 and 1412?
A: For clone 1412: This clone specifically recognizes cytoplasmic domain of CD20 and thus can only be used for intracellular flow cytometry. In this instance you will need to include the fixation and permeabilization steps. Please follow the intracellular flow cytometry staining protocol.

For clone 2H7: This clone is ok for regular surface staining for CD20 and there is no need for any fixation and permeabilization steps.

Our technical protocols can be found here.