IgG1 Negative Control

IgG1 Negative Control

• Background: This ready to use Negative Control reagent contains purified non-conjugated mouse immunoglobulin molecules of IgG1 isotype, which have been selected on the basis of their binding characteristics: no specific binding to Human cell surface or intracellular antigens, same low range of nonspecific binding to Human leukocytes as other Nordic-MUbio Reagents. This isotype control IgG1 is suitable as a Negative Control to be used in combination with Nordic-MUbio reagents for the: - Enumeration of Myeloid Cells - Analysis of Myeloid Differentiation Stage - Enumeration of B-cells and Precursors - Enumeration of T-cells and Precursors - Analysis of Leukemia Cells - Analysis of Immunodeficiency States The negative control reagent 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.
• Host: Mouse
• Isotype: IgG1
• Clone: VI-AP
• Type: Negative Control
• Applications: Direct IF, IF, Flow Cytometry
• Field of Research: Immunology
• Assay Principle: Indirect Immunofluorescence (Staining Procedure) Nordic-MUbio 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 - Incubate the tube for 15 minutes at 4°C or at room temperature in the dak - Add 20 µL of the appropriate conjµgated and diluted secondary antibody - Incubate the tube for 15 minutes at 4°C or at room temperature in te 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 - Centrifµge 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. _x000D_ For “No-Wash” protocol please refer to www.nordicmubio.com Proposed staining procedure for MNC in short: - Carefully add 20 µL antibody conjµgate 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 - Centrifµge tubes for 5 minutes at 300 g - Remove supernatant, resuspend cells in 2-5 mL of phosphate buffered saline (PBS) and centrifµge cells again for 5 minutes at 300 g - Add 20 µL of the appropriate conjµgated and diluted secondary antibody - Incubate the tube for 15 minutes at 4°C or at room temperature in th dark - 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
• Form: 1 mL of; Non-conjµgated VI-AP in PBS pH 7,2, 1% BSA, and 0,05% NaN3, approximately 100 tests
• Precautions: 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.
• Shipping Conditions: Ambient Temperature
• Storage Conditions: 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. If a slight precipitation occurs upon storage, this should be removed by centrifugation; It will not affect the performance or the concentration of the product 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.