Ab181112

In this assessment, we aimed to compare the protein level of each GINS member between pancreatic cancer tissues and adjacent normal tissues. Also, the efficiency of gene transfection was detected. Proteins from pancreatic cancer tissues, adjacent normal tissues, and those correspondingly processed pancreatic cancer cells were extracted using the RIPA lysate. The protein concentration was determined via the BCA assay. Briefly, an equal amount of protein was loaded onto a sodium dodecyl sulfate (SDS) gel. Then, protein bands on the gel were transferred onto a polyvinylidene difluoride membrane, the membrane was blocked for 2 hours using 5% skim milk powder at room temperature. Thereafter, we incubated the membrane for 24 hours at 4°C with primary antibody for GINS1 (ab181112, 1:10,000, Abcam), GINS2 (ab197123, 1:1000, Abcam), GINS3 (ab177515, 1:2000, Abcam), GINS4 (ab139683, 1:1000, Abcam), respectively. Subsequently, the membrane was incubated for 1h at room temperature (RT) with secondary antibodies of the same species. Notably, GAPDH was used as the house-keeping gene. The blotting patterns were detected using ECL (Millipore) and visualized under an X-ray film.