On January 26, 2016Adam Munhall, VA Portland Health Care System wrote:
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Using companies own control lysate (mouse pancreas, 30ug/lane) and our rat brain lysate (20ug/lane) made in RIPA lysis, this antibody (Lot I2010) did label properly at 44-45kDa. Moreover it labeled serum containing expressed Kir6.2 recombinant protein (kindly donated by Dr Show Ling Shyng) at same 44-45kDa location. Lysates were run on Bio-Rad precast gels, wet transferred to Amersham protran membrane, primary dilution 1:200, fluoresced with DyLight800-4Xpeg secondary (1:5000-10000) from Thermo-Pierce, and imaged on a Licor Odyssey. There are many non-specific bands that also appear in our lysate, and around 75kDa in the SCB pancreas control lysate, so giving it 4 of 5 stars. Also, 1:200 is required unless Kir6.2 is very concentrated.
On January 26, 2016 Adam Munhall, VA Portland Health Care System wrote:
Using companies own control lysate (mouse pancreas, 30ug/lane) and our rat brain lysate (20ug/lane) made in RIPA lysis, this antibody (Lot I2010) did label properly at 44-45kDa. Moreover it labeled serum containing expressed Kir6.2 recombinant protein (kindly donated by Dr Show Ling Shyng) at same 44-45kDa location. Lysates were run on Bio-Rad precast gels, wet transferred to Amersham protran membrane, primary dilution 1:200, fluoresced with DyLight800-4Xpeg secondary (1:5000-10000) from Thermo-Pierce, and imaged on a Licor Odyssey. There are many non-specific bands that also appear in our lysate, and around 75kDa in the SCB pancreas control lysate, so giving it 4 of 5 stars. Also, 1:200 is required unless Kir6.2 is very concentrated.