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Invitrogen
Description: The 30-F11 monoclonal antibody reacts with all isoforms of mouse CD45, also known as Leukocyte Common Antigen (LCA). CD45 is expressed by all hematopoietic cells excluding mature erythrocytes and platelets. The cytoplasmic portion of CD45 has tyrosine phosphatase enzymatic activity and plays an important role in activation of lymphocytes.
Applications Reported: This 30-F11 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This 30-F11 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. 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.
CD45 (LCA, leukocyte common antigen) is a receptor-type protein tyrosine phosphatase (PTP) ubiquitously expressed in all nucleated hematopoietic cells, comprising approximately 10% of all surface proteins in lymphocytes. CD45 is absent on non-hematopoietic cell lines, normal and malignant, non-hematopoietic tissues. CD45 glycoprotein is crucial in lymphocyte development and antigen signaling, serving as an important regulator of Src-family kinases. CD45 protein exists as multiple isoforms as a result of alternative splicing, differ in their extracellular domains but share identical transmembrane and cytoplasmic domains. CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells. CD45 isoforms differ in their ability to translocate into the glycosphingolipid-enriched membrane domains and their expression depends on cell type and physiological state of the cell. CD45 has been shown to be an essential regulator of T- and B-cell antigen receptor signaling and suppresses JAK kinases to regulate cytokine receptor signaling. CD45 is also important in promoting cell survival by modulating integrin-mediated signal transduction pathway, DNA fragmentation during apoptosis and inhibition or upregulation of various immunological functions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD45; L-CA; Leukocyte common antigen; Ly-5; Lymphocyte antigen 5; lymphocyte common antigen; Receptor-type tyrosine-protein phosphatase C; T200; T220 and B220
Gene Aliases: B220; Cd45; CD45R; L-CA; loc; Ly-5; Lyt-4; Ptprc; T200
UniProt ID: (Mouse) P06800
Entrez Gene ID: (Mouse) 19264
Molecular Function:
hydrolase
phosphatase
protein phosphatase
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