ELISpot Blue Color Module (Strep-AP and BCIP-NBT), 5 Plate

The complete ELISpot kit is highly sensitive, a microplate-based method for detecting cytokine secreting cells. The kit can be used to detect and enumerate one or two analytes simultaneously. The complete ELISpot kit is ready to run and does not require analytical development or improvement. The enzyme-linked immunosorbent assay module contains the basic components required for enzyme-linked immunosorbent assay.

The ELISpot development module is an alternative to the ELISpot kit. A basic understanding of the development of ELISpot analysis is necessary for the successful use of these reagents. Each researcher should optimize coating conditions, detection sensitivity, types of enzymes and substrates, and the concentration of capture and detection antibodies to achieve the desired results. The analyte-specific ELISpot Development Module and ELISpot Blue Color Module contain components required for analyte detection and visualization, respectively. These modules can be used together, but sold separately. Each module contains enough reagents for at least 5 96-well microplates.

1.An economical alternative to ELISpot Kits.
2.Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time.
3.Generic development protocols provide direction to start an optimization protocol.
4.Customize the assay to your specific needs.

ELISpot

Buffers, Chelators and Reagents

Sandwich

Quantitative Sandwich ELISA Development Module

Whole Cells

1 Pack (for 5 Plates)

1.Streptavidin-AP Concentrate.
2.BCIP/NBT Chromogen.

1.PBS - 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered.
2.Wash Buffer - 0.05% Tween 20 in PBS.
3.Reagent Diluent - 1% BSA in PBS, pH 7.2 - 7.4, 0.2 µm filtered.
4.Deionized H2O.

100 µL

3 hours 35 mins to 4 hours 50 mins**

Pre-coated

96-well microplate, sold separately

The BCIP/NBT Chromogen is highly flammable and toxic if swallowed, in contact with skin, or inhaled.

Store the unopened product at 2 - 8 °C.

Do not use past kit expiration date. This kit is validated for single use only.

Each investigator should optimize the experimental conditions, such as cell type, cell stimulation conditions and cell dilutions of the assay.

For Research Grade Use Only.

1.Calculate the total volume of Streptavidin-AP needed and dilute Streptavidin-AP Concentrate with Reagent Diluent to a working dilution of 1:60.
2.Add 100 µL of the diluted Streptavidin-AP into each well and incubate for 2 hours at room temperature.
3.Wash the plate 3 times with Wash Buffer. Rinse again with deionized water, and remove excess water by inverting the plate and blotting it against a clean paper towel.
4.Add 100 µL of BCIP/NBT solution into each well. Cover the plate and incubate in the dark for 30 minutes at room temperature.
5.Rinse with deionized water. Invert plate and tap to remove excess water and allow the plate to dry at room temperature or at 37°C.
6.Spots can be quantified manually using a dissection microscope or automatically by using a specialized automated ELISpot reader.