Team:UCLA/Notebook/Honeybee Silk/4 August 2015

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Running Tests on Purified Honeybee Protein

• As per Michael's suggestion I am running a Dry Film Test in order to determine the weight by volume percent of silk protein in our samples.
• Tristan is running a BCA assay with Jessica on the samples from 5/26 and 7/29.

Dry Film Test

• Weighed out 100 ul of 5/26, 7/29 and a 3% SDS solution as a control.
• 3%SDS = 0.0991 g, 5/26 silk = 0.1001 g, 7/29 silk = 0.0940 g
• Need to keep in mind that the 5/26 ad 7/29 silks are not directly comparable, because one was solubilized in 750 ul of 3% SDS and the 5/26 was solubilized in 2 ml of 3% SDS, and the 5/26 was from a 250 ml culture, and the 7/29 was from a 100 ml bacterial culture.

BCA

• Using a Thermo Kit, Jessica prepared 9 standards.
• I made a working reagent of 25ml reagent A mixed with 0.5ml reagent B by inverting.
• 100ul of each standard, sample, and blank was added to microcentrifuge tubes.
• 1ml of the working reagent was added to each tube, except the blank, and vortexed for 10s.
• All tubes were incubated at 37C for 45min.
• All samples were transferred to cuvets and run in a spectrophotometer.