ELISpot Path: Vaccinia Human IFN-γ PRODUCT CODE: 3420-4AST-P7-1 CONTENTS: 1 pre-coated strip plate (mAb 1-D1K) and empty plate frame Detection mAb: 7-B6-1, biotin, 1 mg/ml (40 μl) Streptavidin-ALP (40 μl) PepPool: Vaccinia (CD4), human (1 vial) PepPool: Vaccinia (CD8), human (1 vial) Positive control: Anti-CD3 mAb (CD3-2) (100 μl) BCIP/NBT-plus substrate (25 ml) The detection antibody is supplied in sterile filtered (0.2μm) PBS with 0.02% sodium azide. Streptavidin-ALP is supplied in 0.1 M Tris buffer with 0.002% Kathon CG. Anti-CD3 mAb is supplied in sterile filtered (0.2 μm) PBS. Vials have been overfilled to ensure recovery of the specified amount. STORAGE: Shipped at ambient temperature. On arrival all reagents should be stored at 4-8 °C except the peptide pools that should be stored frozen at -20 °C or below. Plates may be kept at room temperature. The expiry date indicates how long unopened products, stored according to instructions, are recommended for

PLEASE READ THROUGH BEFORE STARTING THE ASSAY A Preparation of ELISpot plate (sterile conditions) 1. Assemble the required number of strips in the extra plate frame and wash 4 times with sterile PBS (200 μl/well). Seal the bag with the remaining strips and store at room temperature. 2. Condition the plate with medium (200 μl/well) containing 10% of the same serum as used for the cell suspensions. Incubate for at least 30 minutes at room temperature. B Incubation of cells in plate (sterile conditions) 1. The peptide pools can be used separately or mixed. If used separately, dissolve the peptide...

Hints and Comments These suggestions are based on the detection of antigen-specific immune responses using PBMC. If using clones, mixtures of separated cell fractions etc., other protocols may have to be considered. Plate washing Washing of plates can be done using a multi-channel micropipette. In washing steps not requiring sterile conditions (C1-C5), a regular ELISA plate washer can also be used, provided that the washing head is adapted to the ELISpot plates. Cells Both freshly prepared and cryopreserved cells may be used in the assay. However it is recommended that the latter are rested for...

Updated on 2023/11/02 Developed and manufactured by MABTECH AB, Sweden, whose quality management system complies with the standards ISO 9001:2015 & ISO 13485:2016. Quality management systems Medical devices Quality management systems The products are for research use only. MABTECH shall not be liable for the use or handling of the product or for consequential, special, indirect or incidental damages there from. Mabtech AB (Head Office) Sweden Tel: +46 8 716 27 00 [email protected]