T Cell Granzyme B Detection Dot-ELISA Kit

Sample

Cell

Species Reactivity

Human

Detection Method

DotELISA

Intended Use

The T cell Dot-ELISA Kit is one of the most sensitive tests to monitor ex vivo cellular immune responses at single cell level. The assay can accurately detect secreted proteins, such as cytokines, released by e.g. T cells in response to antigen. The cell suspensions, used in the assay, can originate from blood (PBMC), lymphoid, spleen, bone marrow or CNS tissue.

Contents of Kit

1. Coating antibody* 1 vial 4 °C
2. Biotinylated detection antibody* 1 vial 4 °C
3. Streptavidin-HRP conjugate* 1 vial ≤ -20 °C***
4. AEC Stock solution 1 vial (4 ml) ≤ -20 °C***
5. Substrate buffer (10x) 1 vial (2.5 ml) 4 °C
6. Blocking stock solution (10x) 1 vial (4 ml) 4 °C
7. Dilution buffer R (10x)**** 1 vial (4 ml) 4 °C
8. Tween-20**** 1 vial (5 ml) RT***
9. 96-well plate** with lid 2 plates RT
RT Room temperature (temperature between 20 °C and 26 °C)
* Lyophilized
** PVDF membrane-bottomed Millipore plates
*** Store protected from light

Storage

Coating and detection antibodies
The vials with lyophilized coating and biotinylated detection 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 at 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).

Conjugate
The vial with lyophilized streptavidin-HRP conjugate is stable until the expiry date (indicated on the vial) when stored protected from light at ≤-20 °C. After reconstitution, the reagent is stable at 4°C for at least 2 months when kept sterile and protected from light. However, it is strongly recommended that the solution be divided into small aliquots for single use. These aliquots should be stored protected from light at ≤-20 °C (stable for at least one year).

AEC
The AEC stock solution should be stored at ≤-20 °C and is stable until the expiry date (indicated on the vial)*. Tightly close the vial after use. It is recommended that the solution be divided into small aliquots for single use in polypropylene vials. These aliquots should be stored at ≤-20 °C protected from light (stable for at least one year). * Avoid exposure to light and air: tightly close the vial after use. Avoid contact with polystyrene pipettes and vials.

Substrate buffer
The substrate buffer is stable until the expiry date (indicated on the vial) when stored at 4 °C. Tightly close the vial after use.

Blocking and Dilution buffer
The vials with Blocking stock solution and Dilution buffer 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 T cell Dot-ELISA Kit is one of the most sensitive tests to monitor ex vivo cellular immune responses at single cell level. The assay can accurately detect secreted proteins, such as cytokines, released by e.g. T cells in response to antigen. The cell suspensions, used in the assay, can originate from blood (PBMC), lymphoid, spleen, bone marrow or CNS tissue.