Author:Dang Vu Hoang Source:Laboratory of Vet.Biochemistry and Molecular Biology, Chungbuk National University, Republic of Korea Abstract: This protocol was used for tissues in detail to detect signals of protein
Preparation of Sample
1. Protein extraction
-Preparation of Tissue: 10-20 mg
-Homogenize sample in 0.6 ml PRO-PREP solution by using homogenizer.
-Incubate at –20℃ for overnight.
-Centrifuge at 14.000 rpm for 10 mins at 40C and transfer supernatant to a fresh tube (1.5 ml tube).
2. Protein measurement
-Prepare sample solution: 50 ml
2 ml Protein plus 48 ml DW (dilution factor: 2)
-Prepare 1X Pro-Measure solution: Dilute 1 part of 10X Pro-Measure solution by adding 9 parts of DW. Vortex sufficiently.
-Add 500 ml 1X Pro-Measure solution into 50 ml sample solution. Vortex sufficiently.
-Incubate for 2-5 mins at RT.
-Measure absorbance at 595 nm (500 ml).
-Adjust Protein concentration: 6 mg/ml.
-Re-measure.
