Description

Kit Content:
1, 20xBinding Buffer(20xBB) 10 ml.
2, 100x Binding Enhancer (100xBE) 2 ml.
Note: Milk powder / BSA Powder is not include in the kit.
Calculation: Make 10 ml 1x Binding Buffer (1xBB) -- mix 20xBB 0.5 ml, 100xBE 0.1 ml, Distilled Water (ml) 9.4 ml and
Milk / BSA 0.5g. please make sure Milk powder / BSA is dissolved completely.
Quick Protocol (For the primary & Second antibody to be used together)
1. The ready Western membrane is blocked in wash buffer contained 5% Milk/BAS, to be shaking at RT for 2 minutes
(5-10 min. are allowed).
2. Put the blocked Membrane in the 1x BB contained the primary and HRP conjugated second antibody together, shake at RT
for 20 minutes, (30-60 minutes is allowed if protein has low expression)
3. Rinse the membrane with wash buffer for 4-6 times and continue to wash it with wash buffer at RT, shake for 2 minutes by
4 times.
4. The membrane has been ready to be developed with ECL.
One Hour Protocol (For the primary & Second antibody to be used separately)
1.The ready Western membrane is blocked in wash buffer contained 5% Milk (or BAS), shake at RT for 2-4 minutes
(5-10 min. are allowed).
2. Put the blocked Membrane in the 1xBB contained acceptable primary antibody, to be shaking at RT for 20 minutes, 30-60
minutes is allowed if there is a week expression.
3. Rinse the membrane with wash buffer for 4-6 times and continue to be washed with wash buffer at RT, shake for 2 minutes
by 4 times.
4. Put the membrane in 1xBB with acceptable second antibody; shake at RT for 20 minutes.
5. Rinse the membrane with wash buffer for 4-6 times and continue to wash it with wash buffer at RT, shake for 2 minutes by
4 times.
6. The membrane has been ready to be developed with ECL.

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Antigen-Antibody Binding Enhancer Kit

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