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CD45 Antibody (M005T02B07-A) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 0.4 µg of CD45 Monoclonal Antibody, NovaFluor Blue 660-40S (right). All cells were co-stained with Ter-119 Monoclonal Antibody, eFluor 450 (Product # 48-5921-82). Total viable cells were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
产品信息
M005T02B07-A
产品规格
种属反应
Mouse
宿主/亚型
Rat
/ IgG2b, kappa
分类
Monoclonal
类型
Antibody
克隆号
30-F11
偶联物
激发/发射光谱
508/664 nm
查看光谱
形式
Liquid
浓度
0.1 mg/mL
纯化类型
Affinity chromatography
保存液
PBS, pH 7.2, with BSA
内含物
0.09% sodium azide
保存条件
4°C, store in dark, DO NOT FREEZE!
RRID
产品详细信息
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.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This 30-f11 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This may be used at less than or equal to 0.4 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
Excitation: 492 nm; Emission: 665 nm; Laser: 488 nm (Blue) Laser
靶标信息
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.
仅用于科研。不用于诊断过程。未经明确授权不得转售。
How to use the Panel Builder
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生物信息学
蛋白别名: CD45; L-CA; Leukocyte common antigen; Ly-5; Lymphocyte antigen 5; lymphocyte common antigen; Receptor-type tyrosine-protein phosphatase C; T200; T220 and B220
基因别名: B220; Cd45; CD45R; L-CA; loc; Ly-5; Lyt-4; Ptprc; T200
UniProt ID: (Mouse) P06800
Entrez Gene ID: (Mouse) 19264
activation of MAPK activity
negative regulation of T cell mediated cytotoxicity
positive regulation of T cell mediated cytotoxicity
negative regulation of cytokine-mediated signaling pathway
hematopoietic progenitor cell differentiation
immunoglobulin biosynthetic process
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
negative regulation of protein kinase activity
protein dephosphorylation
negative regulation of cell adhesion involved in substrate-bound cell migration
leukocyte cell-cell adhesion
response to gamma radiation
dephosphorylation
B cell differentiation
T cell differentiation
positive regulation of B cell proliferation
negative regulation of protein autophosphorylation
heterotypic cell-cell adhesion
peptidyl-tyrosine dephosphorylation
T cell proliferation
B cell proliferation
positive regulation of T cell proliferation
positive regulation of MAPK cascade
cell cycle phase transition
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
bone marrow development
stem cell development
negative regulation of peptidyl-tyrosine phosphorylation
T cell receptor signaling pathway
B cell receptor signaling pathway
regulation of 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
regulation of extrinsic apoptotic signaling pathway
positive regulation of extrinsic apoptotic signaling pathway
