Mouse IgG/IgM Dual-Color B Cell FluoroSpot Kit

Capture antibodies specific for Mouse IgG/IgM Dual Color Kits are coated on a 96-well PVDF-backed microplate. Appropriately stimulated cells are added to the microplate which is then placed in a humidified 37 °C CO₂ incubator for the desired period of time. During this incubation period, the immobilized capture antibodies in the immediate vicinity of the secreting cells bind secreted IgG/IgM Dual Color Kits. At the desired time point, one analyte is detected using a biotinylated detection antibody and NorthernLights™ (NL) 557-conjugated to Streptavidin (red flurophore). The second analyte is detected using a NL493-conjugated detection antibody (green fluorophore). Following incubation with the NorthernLights Fluorescence Enhancer, spots form to indicate the presence of IgG/IgM Dual Color Kits secreting cells. Each spot represents an individual secreting cell. The spots can be counted with an ELISpot reader system or using an epifluorescence microscope.

Product Features for Mouse IgG/IgM Dual-Color B Cell FluoroSpot Kit

• Detect and quantitate individual cells secreting IgG/IgM Dual Color Kits
• High sensitivity - ELISpot assays can measure responses with frequencies well below 1 in 100,000 cells
• No in vitro expansion of cells required
• ELISpot assays use only a small number of primary cells

Kit Contents for Mouse IgG/IgM Dual-Color B Cell FluoroSpot Kit

• PVDF-backed Microplate
• Capture Antibody Concentrates
• Biotin-conjugated Detection Antibody
• Streptavidin-conjugated to NL557
• NL493-conjugated Detection Antibody
• Dilution Buffers
• Wash Buffer Concentrate
• NorthernLights Fluorescence Enhancer
• Mouse IgG/IgM Dual Color Kits Positive Controls

Other Reagents Required

• Pipettes and pipette tips
• Deionized or distilled water
• Squirt bottle, manifold dispenser, or automated microplate washer
• 500 mL graduated cylinder
• 37 °C CO₂ incubator
• Sterile culture media
• PBS
• 35% ethanol or methanol
• Block Buffer (1% BSA and 5% sucrose in PBS)
• ELISpot reader capable of detecting fluorescence or a conventional epifluorescence microscope