COMBI Surface: IgG2a Negative Control (FITC) and IgG1 Negative Control (PE)


Catalogue number: GCT-202

Clone4H1-A7 and VI-AP
IsotypeIgG2a and IgG1
Product Type Negative Control
Units50 Tests
Species reactivity Mouse
Application Immunofluorescence
Background
Each staining performed with specific monoclonal antibodies should be paralleled by a staining with an appropriate istotype matched control antibody, in order to be able to control for non-specific binding. The COMBI-REAGENT-Negative Control permits to estimate the degree of non-specific binding of isotype matched immunologbulins to leukocytes via e.g. Fc-receptors. It enables the expert to set flow cytometric parameters accordingly. Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation. Analyses performed with this antibody should be paralleled by positive and negative controls. If unexpected results are obtained, which cannot be attributed to differences in laboratory procedures, please contact us.

Product
PBS pH 7.2, 1% BSA, 0.05% NaN3

Applications
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 4°C 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 2-8°C 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 2-8°C in the dark. Analyze fixed cells within 24 hours

Specificity
The monoclonal antibodies have no known specificity for human leukocytes. It is recommended to use similar amounts of test antibody and isotype matched COMBI-REAGENT Negative Control antibody in each experiment.

Storage
Nordic-MUbio monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8°C (DO NOT FREEZE!) and protected from prolonged exposure to light. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.

Caution
For professional users only. This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.