Protease Inhibitors
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<center> <h1> <b><font>Protease Inhibitors</font> </b></h1> </center>
<center> <font>from Alberts' Lab Protocols (Kathy Miller/Chris Field)</font></center>
Protease inhibitors are rapidly inactivated in aqueous solutions and should be added to solutions immediately before use. Cocktail solutions are stored at -20 degrees and are good for approximately 6 months.
I. Protease Inhibitor Cocktail
The Protease inhibitor cocktail is used as 100X for extract preps and as 1000X for column elution bufferundefined._Cocktail_is_made_in_100~7_EtOH_and_contains~I~K~Hp~M~2~1~Kp~M~2~1~0
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inhbitor |
<center> <font>Sigma catalog #</font></center> |
<center> <font>powder storage</font></center> |
<center> <font>concentration in cocktail</font></center> |
<center> <font>amt for 25 mls cocktail</font></center> |
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Benzamidine HCL |
<center> <font>B6506</font></center> |
<center> <font>4 deg C</font></center> |
<center> <font>1.6 mg/ml</font></center> |
<center> <font>40 mg</font></center> |
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Phenanthroline |
<center> <font>P1294</font></center> |
<center> <font>room temp</font></center> |
<center> <font>1 mg/ml</font></center> |
<center> <font>25 mg</font></center> |
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Aprotiniundefined |
<center> <font>A1153</font></center> |
<center> <font>4 deg C</font></center> |
<center> <font>1 mg/ml</font></center> |
<center> <font>25 mg</font></center> |
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Pepstatin undefined |
<center> <font>P5318</font></center> |
<center> <font>4 deg C</font></center> |
<center> <font>1 mg/ml</font></center> |
<center> <font>25 mg</font></center> |
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Leupeptiundefined |
<center> <font>L2884</font></center> |
<center> <font>-20 deg C</font></center> |
<center> <font>1 mg/ml</font></center> |
<center> <font>25 mg</font></center> |
~undefined_some_components_are_not_completely_soluble_in_100~7_EtOH._Vortex_solution_well_before_use_to_obtain_a_cloudy_suspension_and_add_to_buffer_~F_they_become_soluble_in_aqueous~K~Hp~M~2~1~Kp~M~2~1~0PMSF_is_very_unstable_in_aqueous_solutions._The_100_mM_stock_solution_is_made_in_isopropanol_and_stored_in_the_dark_at_~F20_degrees._Must_be_warmed_~A37_degrees~B_to_get_into_solution._Use_at_1_mM_in_solutions.~K~Hp~M~2~1~Kp~M~2~1~0
compiled by Chad Rappleye
Aroian Lab Protocols
