tcfe4169 Mouse S100A8 (S100 Calcium Binding Protein A8) ELISA Kit

Sample Collection and Storage (universal)

 Serum: Place whole blood sample at room temperature for 2 hours or put it at 4°C overnight and centrifugation for 20 minutes at approximately 1000×g, Collect the supernatant and carry out the assay immediately. Blood collection tubes

should be disposable, non-pyrogenic, and non-endotoxin.

 Plasma: Collect plasma using (EDTA-Na2 or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g at 2 - 8°C within 30 minutes of collection. Collect the supernatant and carry out the assay immediately. Avoid hemolysis, high cholesterol samples.

 Tissue Homogenates: As hemolysis blood has relation to assay result, it is necessary to remove residual blood by washing tissue with pre-cooling PBS buffer (0.01M, pH=7.4). Mince tissue after weighing it and get it homogenized in PBS (the volume depends on the weight of the tissue. Normal, 9mL PBS would be appropriate to 1 gram tissue pieces. Some protease inhibitors are recommended to add into the PBS) with a glass homogenizer on ice. To further break the cells, you can sonicate the suspension with an ultrasonic cell disrupter or subject it to freeze-thaw cycles. The homogenates are then centrifuged for 5minutes at 5000×g to get the supernatant. The total protein concentration was determined by BCA kit and the total protein concentration of each pore sample should not exceed 0.3mg.

 Cell Culture Supernatant: Centrifuge supernatant for 20 minutes at 1000×g at 2 - 8°C to remove insoluble impurity and cell debris. Collect the clear supernatant and carry out the assay immediately.

 Cell Culture Lysate: Commercial RIPA kits are recommended to follow the instructions provided. Generally, 0.5ml RIPA lysis buffer would be appropriate to 2x106 cells, DNA must to be removed. The total protein concentration was

determined by BCA kit and the total protein concentration of each pore sample should not exceed 0.3mg.

 Other Biological Fluids: Centrifuge samples for 20 minutes at 1000×g at 2-8°C. Collect supernatant and carry out the assay immediately.

Note: Samples to be used within 5 days can be stored at 4°C, besides that, samples must be stored at -20°C (assay ≤1 month) or -80°C(assay≤2 months) to avoid loss of bioactivity and contamination. Avoid multiple freeze-thaw cycles. The hemolytic samples are not suitable for this assay.