Mouse anti CD22, conjugated to FITC, IgG1, Clone: [RFB4], Monoclonal
Biozol Catalog Number:
NMB-GM-4052
Supplier Catalog Number:
GM-4052
Alternative Catalog Number:
NMB-GM-4052
Manufacturer:
NordicMubio
Host:
Mouse
Category:
Antikörper
Application:
FC, IF
Species Reactivity:
Human
Conjugation:
FITC
The epitope recognized by antibody RFB4 is a 135 kDa type I integral membrane protein expressed by human B-cells. Precursor B-cells are surface-CD22 negative, but cytoplasmic CD22 positive. Mature B-lymphocytes express CD22 also on their surface. The RFB
2 ml of FITC-conjugated anti CD22 (clone RFB4) in PBS pH 7.2, 1% BSA, and 0.05% NaN3, approximately 100 tests.
Purity:
Purified by Chromatography
Form:
FITC
Formula:
PBS pH 7.2, 1% BSA, 0.05% NaN3
Application Notes:
Staining Procedure for Surface CD22: 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 Nordic-MUbio-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 Staining Procedure for Cytoplasmatic CD22: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM (Cat. No. GAS-002) intracellular CD22 can be easily stained in cell suspensions. - For each sample to be analyzed add 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube - Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature) - Incubate for 15 minutes at room temperature - Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g - Remove supernatant and add to cell pellet 100 µl Reagent B (Permeabilization Medium) and 20 µl of the CD22 monoclonal antibody conjugate - Vortex at low speed for 1-2 seconds - Incubate for 15 minutes at room temperature - Wash cells with phosphate buffered saline as described above - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1.0 % formaldehyde and store them at 2- 8C in the dark. - Analyze fixed cells within 24 hours.
GM-4052
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