|Preparation method :
||Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation and then centrifuged to clarify the lysate. The cell lysate was boiled for 5 minutes in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
|Lysis buffer :
||Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF
|Quality control Testing :
||12.5% SDS-PAGE Stained with Coomassie Blue
|Recommended Usage :
||1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200μL pure water and boiled for 2-5 min.3. Store it at -80°C. Recommend to aliquot the cell lysate into smaller quantities for optimal storage. Avoid repeated freeze-thaw cycles.Notes:The lysate is ready to load on SDS-PAGE for Western blot application. If dissociating conditions are required, add reducing agent prior to heating.
||Samples are stable for up to twelve months from date of receipt at -80°C
|Storage Buffer :
||50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF
|Storage Instruction :
||Lysate samples are stable for 12 months from date of receipt when stored at -80°C. Avoid repeated freeze-thaw cycles. Prior to SDS-PAGE fractionation, boil the lysate for 5 minutes.