![]() Not intended for human, animal diagnostic or therapeutic uses. The apparent molecular weight (kDa) of each protein has been determined by calibration against unstained protein standards supplemental data should be considered for more accurate adjustment in different electrophoresis conditions. Additional data should be considered for a more accurate adjustment.Īpproximately 0.1~0.4 mg/ml of each protein in the buffer (20 mM Tris-phosphate, pH 7.5 at 25☌), 2 % SDS, 0.2 mM Dithiothreitol, 3.6 M Urea, and 15 % (v/v) Glycerol. Note: The molecular weight of each protein (kDa) was measured against an unstained protein ladder in every electrophoresis condition. Precision Plus Protein WesternC Blotting Standards, 1.25 ml. In addition to providing visual verification of transfer at every step, stain-free imaging enables total protein normalization in each lane. Visualize total protein on gels and blots using Bio-Rad’s Stain-Free Gels featuring our proprietary Stain-Free Technology. The entire western blotting workflow consists of several individual steps, each of which is. To verify protein transfer, stain the membrane with Ponceau S after blotting. Load equal amounts of protein (20 g) into the wells of a mini (8.6 x 6.7 cm) or midi (13.3 x 8.7 cm) format SDS-PAGE gel, along with molecular weight markers. All from a starting mixture of proteins extracted from cells or tissues. Life Science Group General Protocol for Western Blotting Protein separation by gel electrophoresis 1. 2.5 μl per well for general Western transferring. Western blotting is a powerful technique that allows you to positively detect your proteins, estimate quantities, and determine their molecular weights. Refer to the Blu13 (BLUelf) Prestained Protein Ladder patterns in various electrophoresis conditions:ģ μl or 5 μl per loading for clear visualization during electrophoresis on 15-well or 10-well mini-gel, respectively. Guide for Molecular Weight Estimation (kDa) The Blu13 Prestained Protein Ladder (BLUelf) keeps track of the size and separation of proteins during SDS-polyacrylamide gel electrophoresis, approximating protein size and validating Western transfer efficiency on PVDF, nylon, or nitrocellulose membranes. The 13 recombinant proteins are covalently coupled with blue chromophore, while 1 green band at 25 kDa and a red band at 75 kDa serve as reference bands. The Blu13 Prestained Protein Ladder (BLUelf) is a combination of 13 pre-stained proteins with molecular weights from 5 to 245 kDa. ![]()
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