Human bcl 2 elisa kit

In brief, all prostae tissue samples were grounded individually in the liquid nitrogen, and then lysed by BioPlex lysis buffer (Bio-Rad, Hercules, CA) inside the microcentrifuge tubes. The tissue lysate was homogenized in a Dounce homogenizer, followed by centrifugation with 13,000 rpm for 10 min at a temperature of 4 °C to obtain a clear supernatant containing the lysate of prostate tissue.
The Bcl-2 level was quantified in all tissue lysate samples by the Human Bcl-2 ELISA Kit (Abcam) based on the related kit protocol.
The expressin level of anoikis-related proteins including PTPN13, TWIST, N-cadherin and E-cadherin in prostate tissue lysates were measured using Human Tyrosine-Protein Phosphatase Non-Receptor Type 13 (PTPN13) ELISA Kit (MyBioSource, USA), TWIST ELISA Kit (Aviva Systems Biology, CA, USA) and Human E-Cadherin, N-Cadherin ELISA Kit (Abcam, Cambridge, MA, USA) respectively, based on the related kit protocols.

The intracellular content of Bcl-2 was quantified using the human Bcl-2 ELISA kit (Abcam, Cambridge, MA, USA). For the sample preparation, the cells were lysed with 1X lysis buffer. After 1 h of incubation at room temperature, the sample was spun at 1,000 g for 15 min and the supernatant was used for Bcl-2 measurement. Briefly, 80 μL of sample diluent and 20 μL of sample were added into the wells coated with monoclonal antibody to human Bcl-2, and after that the wells were washed twice with the wash buffer. Next, 50 μL of biotin-conjugate were added into the wells. After 2 h of incubation at room temperature on a microplate shaker, 100 μL of streptavidin-horseradish peroxidase were added. The sample was incubated again for 1 h at room temperature on a microplate shaker. The solution was withdrawn and the wells were washed with 3X wash buffer. Immediately, 100 μL of TMB substrate solution were added. Finally, 100 μL of stop solution were added into each well to stop the enzyme reaction. The absorbance was read using an ELISA reader at 450 and 620 nm wavelengths. Bcl-2 protein concentration was determined from the standard protein graphic using the optical densities of the samples^{32 (link)}.