PBSTComponents50mMsodium.PDF

PBST Components: 50 mM sodium phosphate buffer (Sigma-Aldrich) (pH 7.5) containing 155 mM NaCl (Sigma- Aldrich)and Tween? 20 (Sigma-Aldrich) 0.05%. Troubleshooting background issues when Detection procedure after protein electrophoresis and using the iBlot? Dry Blotting System blotting (with either the iBlot? Dry Blotting System or the XCell II? Blot Module) using the WesternBreeze? Galia Feldman2, Inbal Simcha2, Reut Mali2, Cory Szafranski1, Chemiluminescent Immunodetection Protocol: and Heath Balcer1 1. Block membrane for 30 minutes with 10 ml blocking 1Invitrogen Corporation, Carlsbad, CA, USA solution (TBST/PBST + 0.1% Tween? 20 + Skim Milk 5%, or 2Invitrogen Corporation, Ness Ziona, Israel WB blocking solution). 2. Wash membrane in 20 ml water for 5 minutes. Introduction 3. Incubate with 10 ml primary ab 1 hour + Blocking solution. 4. Wash membrane in 20 ml PBST/TBST/WB wash 3 x 5 minutes. A frequent problem that arises when analyzing western 5. Incubate 30 min in 10 ml