![]() ![]() Incubate for 2 hr at RT, or overnight at 4☌. Rinse the blot briefly with wash buffer and then add primary antibody diluted in the wash buffer (a concentration of 1-10 µg/ml is generally acceptable, but check datasheets for precise recommendations). Place blot into blocking solution for 2 hr at RT, or overnight at 4☌. ![]() PBS Disodium potassium phosphate, 1.15g Distilled water, 1 L Potassium chloride, 0.2 g Potassium dihydrogen phosphate, 0.2g Sodium chloride, 8.0 g PBST Disodium potassium phosphate, 1.15 g Distilled water, 1 L Potassium chloride, 0.2 g Potassium dihydrogen phosphate, 0.2g Sodium chloride, 8.0 gįollowing SDS-PAGE, transfer proteins onto blotting membrane according to the manufacturer’s instructions.Ĭheck protein transfer by staining the blot with Ponceau S for 1 min, then completely destain the blot by washing with distilled water. Washing buffer Blocking buffer + 0.1% Tween 20 Ponceau S Acetic acid, 5 ml However, we advise using our protocol for detection of phosphorylated proteins by western blot. *Note: For cleaner western blots, Block Ace is recommended over 5% non-fat dried milk dissolved in PBS. For the detection of phosphorylated protein, use the recommended blocking solution as stated on the product datasheet. The Immun-Star WesternC kit allows researchers to take full advantage of the features offered by their imaging instruments.Blocking Buffer Block Ace BUF029 dissolved in water, or 5% non-fat dried milk dissolved in PBS. The Immun-Star WesternC kit generates an intense signal immediately that is easily captured and repeatedly imaged by CCD cameras such as Bio-Rad's Molecular Imager VersaDoc and ChemiDoc XRS Systems. To utilise and complement the Precision Plus Protein WesternC standards, the Immun-Star WesternC kit is specifically designed for CCD imagers. ![]() Optimized for CCD imagers, the Precision Plus Protein WesternC standards are the sharpest in the industry and ideal for visualization of blots, production of publication-quality images of bands on blots, and accurate molecular weight estimation direction from acquired blot images. The 25, 50 and 75kD bands are stained pink for easy band reference and blot orientation, with all other bands prestained blue.Įach standard has an integral Strep-tag sequence to enable parallel detection on western blots using StrepTactin-HRP conjugates, also provided by Bio-Rad. The Precision Plus Protein WesternC standard offers ten prestained protein bands over the same broad range as all Precision Plus Protein standards, ie, from 10 to 250kD. ![]() This allows users to monitor electrophoresis progress, assess transfer efficiency and direct visualisation of the WesternC standards through Western blotting. The Precision Plus Protein WesternC Standard.Īpplicable for all 1D gel electrophoresis experiments, the Precision Plus Protein WesternC Standard combines the convenience of prestained bands with western blot detection. The complementing Immun-Star WesternC kit will further establish Bio-Rad as a leading provider of protein standards and allow users to visualise the standards on CCD imagers, it says. The Immun-Star WesternC kit is a complementary chemiluminescent detection kit that is optimised for CCD imaging of the WesternC standards and protein samples.Ĭurrently, Bio-Rad offers a sample size of both the WesternC standard and the Immun-Star WesternC kit.īio-Rad envisions that the Precision Plus Protein WesternC standard will become the leading western blotting standard in all areas of biological research. The Precision Plus Protein WesternC standards offers ten prestained protein bands that can also be detected through chemiluminescent western blotting. Bio-Rad Laboratories has announced the release of a broad-range, prestained chemiluminescent protein standard and a chemiluminescent detection kit for protein sizing and expression analysis. ![]()
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