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Prep Bradford Reagent, Turn on spectrophotometer - Coggle Diagram
Prep Bradford Reagent
+50mL 95% Ethanol
+100mg Coomassie Brilliant Blue G-250, dissolve it
+100mL 85% Phosphoric Acid
When dye completely dissolved, dilute to 1L
Filter thru Whatman#1 JUST BEFORE USE
Turn on spectrophotometer
Set to 595 nm wavelength
Prep standard Curve
Get 5 1.5mL microcentrifuge tubes
+aliquots of 1.0mg/mL BSA
(5.0, 10.0, 25.0, 50.0, 75.0 uL)
Each test tube, bring vol to total of 100uL w/ 0.15M NaCl
Get separate test tube
+100uL of 0.15M NaCl
+1mL Bradford Reagent to EACH test tube
Vortex it
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Prepare unknown
Get 1.5mL microcentrifuge tube
+1mL Bradford Regent to 100uL of unknown protein
Vortex
Leave for 2 mins at room temp
Measure the Abs595 protein concetration
Use standard curve to compute concentration
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