IFN-γ/IL-4 Detection Dot-ELISA Kit

Sample

Cell

Species Reactivity

Human

Detection Method

DotELISA

Intended Use

The dual-color dot-ELISA kit assay is a modification of the T cell ELISPOT assay, one of the most sensitive assays for monitoring ex vivo cellular immune responses at the single cell level. The dot-ELISA kit was developed for the simultaneous detection of two secreted proteins, such as cytokines, which are released by e.g. T cells in response to an antigen. The cell suspensions, used in the assay are usually derived from blood (PBMC).

Contents of Kit

1. Coating antibodies* 2 vials 4 °C
2. Detection antibodies* 2 vials 4 °C**
3. Fluorescent-labeled conjugate* 1 vial 4 °C**
4. Blocking stock solution (10x) 1 vial (4 ml) 4 °C
5. Dilution buffer R (10x) 1 vial (4 ml) 4 °C
6. Tween-20 1 vial (5 ml) RT**
7. Spot enhancer (4x) 1 vial (6 ml) 4 °C
8. 96-well FluoroSpot plate*** with lid 2 plates RT
9. Adhesive cover slip 5 slips RT
RT Room temperature (temperature between 20 °C and 26 °C)
* Lyophilized
** Store protected from light
*** PVDF membrane-bottomed plates with low background fluorescence

Storage

Coating antibodies
The vials with lyophilized coating antibody can be safely stored at 4 °C until the expiry date (indicated on the vials). After reconstitution, the antibodies are stable at 4 °C for at least 12 months when kept sterile. However, it is recommended that the reconstituted antibody solutions be divided into small aliquots for single use. These aliquots should be stored at ≤-20 °C (stable for at least two years).
Detection antibodies
The vials with lyophilized detection antibody are stable until the expiry date (indicated on the vials) when stored protected from light at 4 °C. After reconstitution, the antibodies are stable at 4 °C for at least 12 months when kept sterile and protected from light. However, it is recommended that the reconstituted antibody solutions be divided into small aliquots for single use. These aliquots should be stored protected from light at ≤-20 °C (stable for at least two years).
Conjugate
The vial with lyophilized fluorescent-labeled conjugate is stable until the expiry date (indicated on the vial) when stored at 4 °C protected from light. After reconstitution, the reagent is stable at 4 °C for at least 6 months when kept sterile and protected from light.
DO NOT FREEZE.
Blocking stock solution, Dilution buffer and Spot enhancer. The vials with Blocking stock solution, Dilution buffer and Spot enhancer can be safely stored at 4 °C until the expiry date (indicated on the vial). After opening, these solutions are stable for at least 6 months when kept sterile.
Tween-20
Tween-20 can safely be stored at RT (protected from light) and is stable until the expiry date (indicated on the vial).

General Notes

The dual-color dot-ELISA kit assay is a modification of the T cell ELISPOT assay, one of the most sensitive assays for monitoring ex vivo cellular immune responses at the single cell level. The dot-ELISA kit was developed for the simultaneous detection of two secreted proteins, such as cytokines, which are released by e.g. T cells in response to an antigen. The cell suspensions, used in the assay are usually derived from blood (PBMC).