DSP30 (also known as KLH [keyhole limpet hemocyanin] conjugated to a 30-mer oligonucleotide) is a protein/DNA complex that is known to induce proliferation of B cells and can be used in the cytogenetic analysis of B-cell malignancies. The addition of interleukin-2 (IL-2) can enhance the mitogenic effect. However, the optimal concentrations and volumes can vary depending on the specific cell type, the phase of the cell cycle, and the overall health of the cells. Here are some general guidelines:

Concentration of DSP30:

When ordering DSP30 for synthesis, a concentration of around 1-10 μg/mL is often used. However, this can vary, and it is essential to determine the optimal concentration for your specific cell type through preliminary experiments.

The concentration you should order will depend on the volume you plan to use and the number of experiments you intend to conduct. For example, if you want to prepare 10 mL of a 1 μg/mL solution, you would need 10 μg of DSP30.

Volume of DSP30 to Add Per mL of Culture:

Typically, a volume of 0.1-1.0 mL of DSP30 solution is added per mL of cell culture. The exact volume will depend on the concentration of your DSP30 stock solution and the desired final concentration in the culture.

For example, if you have a 1 μg/mL stock solution and you want to achieve a final concentration of 0.5 μg/mL in a 1 mL cell culture, you would add 0.5 mL of the stock solution.

Concentration of IL-2:

The concentration of IL-2 can range from 50 to 200 IU/mL, depending on the protocol and the cell type.

It’s important to optimize the concentration of IL-2 in combination with DSP30 for your specific cell line or sample.

Protocol Example:

Prepare a stock solution of DSP30 at a concentration of 1-10 μg/mL in an appropriate buffer (e.g., PBS).

Add a certain volume of this stock solution to your cell culture to achieve the desired final concentration. For example, add 0.5 mL of a 1 μg/mL stock solution to 1 mL of cell culture to achieve a final concentration of 0.5 μg/mL.

Add IL-2 to the culture at a concentration of 50-200 IU/mL.

Incubate the cells with DSP30 and IL-2 for a period that is optimized for your cell type (commonly 24-72 hours) to allow for cell proliferation and the appearance of metaphases.

Remember that these are general guidelines, and you should perform optimization experiments to determine the best conditions for your specific B cell malignancy samples. It’s also crucial to include appropriate controls in your experiments to ensure that the mitogens are working as expected. Always refer to the literature and established protocols for your specific type of B cell malignancy for the most accurate and up-to-date guidance.