Description: The RA3-6B2 monoclonal antibody reacts with exon A-restricted isoform of mouse CD45, a 220 kDa surface molecule. CD45R/B220 epitope is mainly expressed by the B cell lineage from early Pro-B to mature B cells. However, some activated T cells, lymphokine activated killer cells (LAK), NK cell progenitors in the bone marrow, and T cells of the lpr/lpr mutant mouse also express this antigen.
Applications Reported: This RA3-6B2 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This RA3-6B2 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
CD45R is a member of the protein tyrosine phosphatase (PTP) family. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. CD45R contains an extracellular domain, a single transmembrane segment and two tandem intracytoplasmic catalytic domains. It is specifically expressed in hematopoietic cells and has been shown to be an essential regulator of T- and B-cell antigen receptor signaling.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD45; CD45 antigen; L-CA; Leukocyte common antigen; Ly-5; Lymphocyte antigen 5; lymphocyte common antigen; protein tyrosine phosphatase, receptor type, C; protein tyrosine phosphatase, receptor type, c polypeptide; Receptor-type tyrosine-protein phosphatase C; T200; T200 glycoprotein; T200 leukocyte common antigen
Gene Aliases: B220; CD45; CD45R; GP180; L-CA; LCA; loc; Ly-5; LY5; Lyt-4; PTPRC; T200