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Rencojtres refer to primary antibody datasheet or product webpage for recommended antibody dilution. Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature. Prepare solutions with reverse osmosis deionized RODI or equivalently purified water. Primary Antibody Dilution Buffer: Volumes are for 10 cm x 10 cm cm2 of membrane; for different sized membranes, adjust volumes accordingly.