FluoroSpot Plus: Mouse IFN-γ/IL-2/TNF-α
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FluoroSpot Plus: Mouse IFN-γ/IL-2/TNF-α FSP-414245-2 FSP-414245-10 www.mabtech.com 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 therefrom.

INTRODUCTION FluoroSpot – General description This triple FluoroSpot kit enables convenient enumeration of cells secreting either one, two or three analytes. FluoroSpot combines the sensitivity of ELISpot with the capacity to analyze secretion of several analytes simultaneously. This highly sensitive cellular assay is robust, easy to perform, and suitable both for testing single samples and for large-scale screening. TAG Detection mAb-tag Analyte 1 Capture mAb TAG Detection mAb-biotin Analyte 2 Capture mAb Detection mAb-tag Analyte 3 Capture mAb Our FluoroSpot Plus kits include pre-coated...

FluoroSpot Plus: Mouse IFN-y/IL-2/TNF-a The kit is shipped at ambient temperature. All reagents should be stored at 4-8°C upon arrival. Plates should be stored at room temperature. The expiry date indicates how long unopened products, stored according to instructions, are recommended to remain in use. Component Concentration 2-plate kit 10-plate kit Pre-coated plates Plates pre-coated with mAbs AN18, 1A12, and 2 10 Anti-CD28 mAb is supplied in sterile filtered (0.2 pm) PBS. The biotinylated antibody contains 0.02% sodium azide. The BAM- and WASP-conjugated antibodies,...

Please consult the table below for recommendations on dilutions and cell incubation time. The reagents should not be diluted in advance. For additional information, read the Tips For A Successful Assay section. Component Dilution Detection mAbs Anti-IFN-y mAb (R4-6A2), BAM 1:200 Fluorophore conjugates Anti-BAM mAb, 490 1:200 Cell incubation time overnight A. Plate preparation (sterile conditions) 1. Remove the plate from the sealed package and wash the plate three times with sterile PBS (200 gl/well). 2. Block/condition the plate by adding cell incubation medium containing 10% fetal calf...

1. Remove the cells by emptying the plate and then wash the plate five times with PBS (200|jl/well). 2. Dilute the detection antibodies in PBS containing 0.1% BSA (PBS-0.1% BSA) according to the table above. If more than one analyte is analyzed, dilute the detection antibodies in the same tube. Add 100 jl/well and incubate for 2 hours at room temperature. 3. Wash the plate five times with PBS (200 gl/well). 4. Dilute the fluorophore conjugates in PBS-0.1% BSA to the concentrations as shown in the table above. If more than one analyte is analyzed, dilute the conjugates in the same tube. Add...

TIPS FOR A SUCCESSFUL ASSAY Buffers PBS for washing and dilution should be filtered (0.2 μm) to remove any particles. We do not recommend the inclusion of Tween or other detergents in the washing and incubation buffers. Washing Plates can be washed using a multi-channel micropipette. A regular plate washer can be used in washing steps when sterile conditions are not required, provided that the washing head is adapted to ELISpot/FluoroSpot plates. Serum Serum should be selected to support cell culture and give low background staining. We recommend the use of fetal calf serum. Alternatively,...

PROTOCOL SUMMARY Start Wash x3 Cell incubation Block/condition plate for 30 min. Add stimuli and cells Incubate at 37°C Sterile conditions Remove cells Wash x5 Detection Add detection antibody mix Incubate 2h at RT Wash x5 Add conjugate mix Incubate 1h at RT Wash x5 Add FluoroSpot enhancer Incubate 15 min. at RT Empty plate Remove underdrain and dry plate Analysis

Updated on 2022/11/29 Developed and manufactured by MABTECH AB, Sweden, whose quality management system complies with the standards ISO 9001:2015 & ISO 13485:2016. The products are for research use only. Quality management systems Medical devices Quality management systems 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 mabtech@mabtech.com