Invitrogen™ CD45R (B220) Monoclonal Antibody (RA3-6B2), eFluor™ 570, eBioscience™

Catalog No.	Quantity
50-112-3594	25 μg

Catalog No. 50-112-3594 Supplier Invitrogen™ Supplier No. 41045280

Description

Rat Monoclonal Antibody

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 microscopy, immunocytochemistry, and immunohistochemical staining of formalin-fixed paraffin embedded or frozen tissue sections. Applications Tested: This RA3-6B2 antibody has been tested by immunohistochemistry of frozen mouse spleen and can be used at less than or equal to 1 μg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Filter Recommendation: When using this eFluor™ 570 antibody conjugate, we recommend a filter that will capture the 570 emission wavelength (for example, Excitation 545/25, 565LP, Emission 605/70). A standard Alexa Fluor™ 555 or TRITC filter is acceptable. Excitation: 555 nm; Emission: 570 nm.

CD45R (PTPRC) 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. CD45 contains an extracellular domain, a single transmembrane segment and two tandem intracytoplasmic catalytic domains. CD45 is specifically expressed in hematopoietic cells and has been shown to be an essential regulator of T- and B-cell antigen receptor signaling. CD45 glycoprotein is crucial in lymphocyte development and antigen signaling, serving as an important regulator of Src-family kinases. CD45 protein exists in multiple isoforms as a result of alternative splicing, and these isoforms differ in their extracellular domains, but share identical transmembrane and cytoplasmic domains. Further, these 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. In addition to its role in immunoreceptor signaling, CD45 is important in promoting cell survival by modulating integrin-mediated signal transduction pathway and is also involved in DNA fragmentation during apoptosis. CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells. Diseases associated with CD45 dysfunction include Severe Combined Immunodeficiency (T-cell negative, B-Cell Negative) and Hepatitis C.

Specifications

• Antigen: CD45R (B220)
• Applications: Immunohistochemistry (Frozen), Immunohistochemistry (Paraffin), Immunocytochemistry
• Classification: Monoclonal
• Clone: RA3-6B2
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 570
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: PTPRC
• Gene Accession No.: P06800, P08575
• Gene Alias: B220; Cd45; CD45 antigen; CD45R; GP180; Lca; L-CA; leucocyte common antigen; leukocyte common antigen; leukocyte common antigen A; leukocyte common antigen B; loc; LY5; Ly-5; Lymphocyte antigen 5; lymphocyte common antigen; Lyt-4; membrane tyrosine phosphatase; protein tyrosine phosphatase receptor type C; protein tyrosine phosphatase, receptor type C; protein tyrosine phosphatase, receptor type, C; protein tyrosine phosphatase, receptor type, c polypeptide; Protein tyrosine phosphatase, receptor-type, c polypeptide; Ptprc; Receptor-type tyrosine-protein phosphatase C; RT7; T200; T200 glycoprotein; T200 leukocyte common antigen
• Gene Symbols: PTPRC
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 25 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 19264, 5788
• Target Species: Human, Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2a κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.