Quick-blotsTM is Ready-to-use PVDF membrane with 11 Rat tissue lysates (20 µg total protein per lane). Quick-blotsTM is a very simple method to identify different proteins from rat tissues.
Protocol:
1. Activate the membrane with Methanol for 1-2 min and then wash the blots with TBST (25 mM Tris-Cl, pH 8.0; 125 mM NaCl; 0.1% Tween 20) buffer thrice to get rid of extra methanol.
2. Block the membrane for 1 hour in 5% skimmed milk ( in TBST) to avoid non specific binding of antibodies.
3. Wash the membranes 2 times in TBST and incubate with primary antibodies prepared in 1% skimmed milk for 2 hr in RT/ over night at 4oC.
4. After primary incubation, wash the membranes for 1 hour in TBST at RT. (Buffer should be changed in 30 min interval)
5. Incubate the membranes in appopriate Secondry antibody for 1 hour.
6. After secondary incubation, wash the membranes for 2 hours (Buffer should be changed in 30 min interval). Then develop the blots with Chemiluminescent Substrate and interpret the results in photographic film.