Invitrogen

CD45R (B220) Monoclonal Antibody (RA3-6B2), NovaFluor™ Yellow 660, eBioscience™

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说明书
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说明书
实验方案
问题与答案
技术支持

产品信息

M011T02Y04-A

应用
建议稀释比
已发表文章

流式细胞分析 (Flow)

1.0 µg/test
-
产品规格

种属反应

Human, Mouse

宿主/亚型

Rat / IgG2a, kappa

分类

Monoclonal

类型

Antibody

克隆号

RA3-6B2

偶联物

NovaFluor™ Yellow 660 NovaFluor™ Yellow 660 NovaFluor™ Yellow 660

激发/发射光谱

550/664 nm 查看光谱 spectra

形式

Liquid

浓度

0.1 mg/mL

纯化类型

Affinity chromatography

保存液

PBS, pH 7.2

内含物

0.09% sodium azide

保存条件

4°C, store in dark, DO NOT FREEZE!

RRID

AB_3098977

产品详细信息

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.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

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 1.0 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.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Our internal testing shows that NovaFluor Yellow 660 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.

Excitation: 550 nm; Emission: 664 nm; Laser: 561 nm (Yellow) Laser

靶标信息

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.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

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生物信息学

蛋白别名: 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; PTPRC; Receptor-type tyrosine-protein phosphatase C; T200; T200 glycoprotein; T200 leukocyte common antigen

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基因别名: B220; CD45; CD45R; GP180; L-CA; LCA; loc; Ly-5; LY5; Lyt-4; PTPRC; T200

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UniProt ID: (Human) P08575, (Mouse) P06800

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Entrez Gene ID: (Human) 5788, (Mouse) 19264

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功能
protein tyrosine phosphatase activity transmembrane receptor protein tyrosine phosphatase activity protein binding protein kinase binding phosphoprotein phosphatase activity heparin binding hydrolase activity phosphatase activity heparan sulfate proteoglycan binding protein phosphatase protein modifying enzyme
参与通路
negative regulation of T cell mediated cytotoxicity negative regulation of cytokine-mediated signaling pathway hematopoietic progenitor cell differentiation immunoglobulin biosynthetic process negative regulation of protein kinase activity protein dephosphorylation negative regulation of cell adhesion involved in substrate-bound cell migration cell surface receptor signaling pathway dephosphorylation T cell differentiation positive regulation of B cell proliferation peptidyl-tyrosine dephosphorylation B cell proliferation positive regulation of T cell proliferation cell cycle phase transition positive regulation of protein kinase activity bone marrow development stem cell development T cell receptor signaling pathway B cell receptor signaling pathway positive regulation of antigen receptor-mediated signaling pathway release of sequestered calcium ion into cytosol defense response to virus regulation of cell cycle positive regulation of hematopoietic stem cell migration positive regulation of stem cell proliferation activation of MAPK activity positive regulation of T cell mediated cytotoxicity positive regulation of T cell mediated immunity regulation of humoral immune response mediated by circulating immunoglobulin positive regulation of humoral immune response mediated by circulating immunoglobulin leukocyte cell-cell adhesion response to gamma radiation B cell differentiation negative regulation of protein autophosphorylation heterotypic cell-cell adhesion T cell proliferation positive regulation of MAPK cascade positive thymic T cell selection negative thymic T cell selection regulation of B cell differentiation positive regulation of T cell differentiation positive regulation of gamma-delta T cell differentiation positive regulation of alpha-beta T cell proliferation positive regulation of isotype switching to IgG isotypes negative regulation of peptidyl-tyrosine phosphorylation regulation of B cell receptor signaling pathway regulation of extrinsic apoptotic signaling pathway positive regulation of extrinsic apoptotic signaling pathway
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