Antibody Eluates

During TSA staining, the antibody eluent was employed for the elution of antibodies from paraffin sections, cell climbing sections, bone tissue sections, and frozen sections. The eluted antibodies were then utilized for further analysis. It is important to ensure accurate and precise elution of the antibodies for reliable and reproducible results. Therefore, the antibody elution process must be carefully optimized to achieve optimal conditions for each type of specimen.

Ready to use without dilution.
To elute the antibody, start by preheating the antibody elution solution to 37°C. Then, add a small amount of the solution to cover the sample and let it sit at room temperature for 3 to 5 minutes. After that, discard the elution solution and pour a sufficient amount of fresh elution solution over the sample. The amount of elution solution should be slightly higher than the tablet, typically around 150-300 μL. Maintain the temperature at 37°C for 10 to 40 minutes, although 20 minutes is usually sufficient. Following the elution step, wash the sample three times with PBS and consider adding fixative briefly to ensure proper fixation.

The effectiveness of antibody elution relies on various factors such as slice thickness, elution temperature and time, and the type of primary antibody used. Generally, wet box treatment at 37°C for 10-40 minutes is suitable for 4-8μm slices. However, for antibodies such as E-cadherin and TuJ1 that bind strongly to the antigen, higher temperatures or longer elution times may be necessary. In case of trouble eluting certain antibodies, reducing the dilution ratio of the first antibody or including them in the final round of TSA staining can be helpful. Proper protective measures must be taken to avoid skin and eye contact with reagents, and any accidental contact should be immediately rinsed off with plenty of water. The viability of reagents may also diminish beyond their expiration date, so it is crucial to use them within the recommended time-frame.

store at 2-8° C for 12 months