Mouse anti CD45, IgG2a, Clone: [VIT200], Monoclonal
Biozol Catalog Number:
NMB-GM-4061
Supplier Catalog Number:
GM-4061
Alternative Catalog Number:
NMB-GM-4061
Manufacturer:
NordicMubio
Host:
Mouse
Category:
Antikörper
Application:
FC, IF
Species Reactivity:
Human
The CD45 molecule is typically expressed at high levels on all hematopoietic cells. CD45 is a major component of the glycocalix of these cells and can be expressed in different isoforms. Antibody VIT200 recognizes a pan CD45 epitope, which is expressed o
GM-4061 can be applied in immunohistochemistry on frozen and parrafin embedded tissues and in flow cytometry. Direct Immunofluorescence (Staining Procedure) Nordic-MUbio fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations. Proposed staining procedure for whole blood in short: - For each sample add 50 µl of EDTA anti-coagulated blood to a 3-5 ml tube - Add 20 µl of the appropriate Nordic-MUbio monoclonal antibody conjugate - Incubate the tube for 15 minutes at 4C or at room temperature in the dark - Add 100 µl NM-LYSE (Cat.No. GAS-003) to each tube and incubate for 10 minutes at room temperature - Add 3-4 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature - Centrifuge tube for 5 minutes at 300 g - Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid - Analyze immediately or store samples at 2-8 C in the dark and analyze within 24 hours For ''No-Wash protocol please refer to www.nordicmubio.com Proposed staining procedure for MNC in short: - Carefully add 20 µl antibody conjugate and 50-100 µl MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-8C or at room temperature - Centrifuge tubes for 5 minutes at 300 g - Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8C in the dark. Analyze fixed cells within 24 hours Indirect Immunofluorescence (Staining Procedure) - Mix 20 µl Nordic-MUbio purified antibody with 50 µl whole blood or MNC suspension - Incubate for 15 minutes at 2-8C - Wash cells with phosphate buffered saline (PBS) - Add to cell pellet 20 µl of affinity purified, fluorochrome labeled F(ab)2 anti mouse Ig antibodies - Incubate for 15 minutes at 2-8C - Wash cells with phosphate buffered saline (PBS) or proceed as described for direct staining
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