Invitrogen

CD45.2 Monoclonal Antibody (104), NovaFluor™ Blue 510, eBioscience™

View all (49) CD45.2 antibodies

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Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

M027T02B01-A

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

0.8 µg/test
-
Product Specifications

Species Reactivity

Mouse

Host/Isotype

Mouse / IgG2a, kappa

Class

Monoclonal

Type

Antibody

Clone

104

Conjugate

NovaFluor™ Blue 510 NovaFluor™ Blue 510 NovaFluor™ Blue 510

Excitation/Emission Max

493/513 nm View spectra spectra

Form

Liquid

Concentration

0.1 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

RRID

AB_3099041 (AB_2921162)

Product Specific Information

Description
The 104 monoclonal antibody reacts with the mouse CD45 molecule, the leukocyte common antigen (LCA) in CD45.2-expressing mouse strains. The strains that express CD45.2 include the most commonly used mouse strains C57BL/6, BALB/c, C58, DBA/1, DBA/2, C3H/He, CBA, 129, A and AKR. CD45.2 is expressed by all leukocytes in these strains.

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

Applications Tested
The 104 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.

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 (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: 493 nm; Emission: 513 nm; Laser: 488 nm (Blue) Laser

Target Information

CD45.2 (LCA, leukocyte common antigen) is a receptor-type protein tyrosine phosphatase ubiquitously expressed in all nucleated hematopoietic cells, comprising approximately 10% of all surface proteins in lymphocytes. CD45.2 glycoprotein is crucial in lymphocyte development and antigen signaling, serving as an important regulator of Src-family kinases. CD45.2 protein exists as multiple isoforms as a result of alternative splicing; these isoforms differ in their extracellular domains, whereas they share identical transmembrane and cytoplasmic domains. These 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. Besides the role in immunoreceptor signaling, CD45.2 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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

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Bioinformatics

Protein Aliases: CD45; CD45 antigen; GP180; L-CA; LCA; Leukocyte common antigen; Ly-5; LY5; Lymphocyte antigen 5; lymphocyte common antigen; Receptor-type tyrosine-protein phosphatase C; T200; T200 glycoprotein

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Gene Aliases: B220; Cd45; CD45R; L-CA; loc; Ly-5; Lyt-4; Ptprc; T200

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UniProt ID: (Mouse) P06800

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Entrez Gene ID: (Mouse) 19264

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Function(s)
phosphoprotein phosphatase activity protein tyrosine phosphatase activity protein binding heparin binding hydrolase activity phosphatase activity protein kinase binding heparan sulfate proteoglycan binding
Process(es)
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
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