B Cells IgG2a Detection Dot-ELISA Kit

Sample

Cell

Species Reactivity

Mouse

Detection Method

DotELISA

Intended Use

The B cell ELISPOT assay determines the number of individual antibody secreting cells (ASC) in single cell suspensions and can identify ex vivo antigen-specific memory B cells. The cell suspensions used may originate from blood (PBMC) or secondary lymphoid organs.

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. Recombinant IL-2* 1 vial ≤ -20 °C
5. R848 (Resiquimod) 1 vial (0.5 ml) ≤-20 °C**
6. AEC coloring system:
6.1. I. AEC stock solution 1 vial (4 ml) ≤-20 °C**
6.2 II. Substrate buffer (10x) 1 vial (5 ml) 4 °C
7. Blocking stock solution (10x) 1 vial (10 ml) 4 °C
8. Dilution buffer R (10x) 1 vial (10 ml) 4 °C
9. Tween-20 1 vial (5 ml) RT**
RT Room temperature (temperature between 20 °C and 26 °C)
* Lyophilized
** 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 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).
IL-2
The vial with lyophilized recombinant IL-2 can be safely stored at ≤-20 °C until the expiry date (indicated on the vial). After reconstitution, the solution should be stored in small aliquots at ≤ -70 °C for single use (stable for at least one year).
R848
The vial with R848 can be safely stored at ≤-20 °C until the expiry date (indicated on the vial). It is strongly recommended to divide the solution into small aliquots for single use. These aliquots should be stored at ≤-20 C protected from light (stable for at least one year).
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 ≤-20 °C protected from light at (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 R 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 B cell ELISPOT assay determines the number of individual antibody secreting cells (ASC) in single cell suspensions and can identify ex vivo antigen-specific memory B cells. The cell suspensions used may originate from blood (PBMC) or secondary lymphoid organs.