ELISpot Blue Color Module (Strep-AP and BCIP-NBT), 5 Plate (CAT#: ITS-0322-P157)

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.