Multiplex western blot system makes it easy to detect multiple proteins simultaneously. Some vendors supply multiplex western cocktail to detect multiple proteins in one membrane without stripping and reprobing. The multiplex blotting apparatus are also available from several suppliers. Optimization of western blot is required for multiplex western blot analysis. References: Multiplex Western Blotting System for the Analysis of Muscular Dystrophy Proteins

alkaline phosphatase substrates is a chemiluminescent substrate for western blotting detection. It can also be used with colorimetric and fluorescent western blot detection system.

BSA based blocking solution: bovine serum albumin (BSA) is one of the most commonly used blocking reagent for western blotting. The concentration of BSA is normally between 1-5% in PBS or TBS with 0.05% Tween-20.

Gelatin blocking solution is suitable for nylone or nitrocellulose membrane.Gelatin as blocking agent in Westerns

non-fat dry milk is the most commonly used general blocking protein for western blot blocking. The component of nonfat dry milk is listed bellow. The issue with milk blocking solution is that it may cross-reacting with high sensitive detection reagents.Component of Nonfat Dry Milk (Merck Index):Β-lactoglobulin, α-lactalglobulin, immunoglobulins, serum albumin, three or more different caseins, phosphoproteins, enzymes, hormones, growth factors, nutrient transporters, disease-resistance factors, entire leukocytes, other proteins, lactose, glucose, galactose, amino sugars, sugar phosphates, neutral and acid oligonucleotides, nucleotide sugars, monosaturated fatty acids, polyunsaturated fatty acids, saturated fatty acids, A, B-6, B-12, D, E, H (biotin), folate, niacin, pantothenic acid, riboflavin, thiamin, calcium, iron, magnesium, phosphorous, potassium, sodium, zinc, copper, manganese, and selenium.

Peroxidase substrates is commonly used for western blotting detection. Chemiluminescent Peroxidase Substrates for Western Blotting (Sigma)