Twist 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.

For tissue homogenization, all fresh tissue samples were weighed and the tissue lysate was prepared according to the manufactures protocol (Invitrogen, CA, USA). Approximately 50 μg of each tissue was excised and washed with ice-cold PBS.
The level of p53, RB, IL-1, IL-6, IL-11, NF-kB, NF-κ PTPN13, E-cadherin, N-cadherin and TWIST was assessed using Abcam’s p53 Simple Step ELISA® Kit (Abcam, Cambridge, MA, USA), Human Retinoblastoma ELISA® kit (Sigma-Aldrich, Saint Louis, USA), Human Retinoblastoma ELISA® kit (Sigma-Aldrich, Saint Louis, USA), Human IL-6 ELISA® Kit, Human IL-1 beta ELISA® Kit, Human IL-11 ELISA® Kit, NF-kB p65 Transcription Factor Assay® Kit (Abcam, Cambridge, MA, USA), Human Tyrosine-Protein Phosphatase Non-Receptor Type 13 (PTPN13) ELISA Kit (MyBiosource, USA), Human E-Cadherin, N-Cadherin ELISA Kit (Abcam, Cambridge, MA, USA).and TWIST ELISA Kit (Aviva Systems Biology, CA, USA).