20190521: Protein quantification

Protein extracts:

1. non-transfected HEK cells –> result: 8,35 mg/ml
2. CNTN1-transfected HEK cells (21.05.19) –> result: 33,11 mg/ml
3. CASPR1-transfected HEK cells (21.05.19) –> result: 23,90 mg/ml
4. TE671 (21.05.19) –> result: 8,06 mg/ml
5. undifferentiated HTB11 cells (23.04.19) –> result: 3,17 mg/ml
6. Brain mouse (11.04.19) –> result: 11,24 mg/ml
7. Sciatic nerve lysate (17.05.19) –> result: 0,92 mg/ml

DC Protein Assay – BioRad-

• Dilute BSA to 4 mg/ml –> stock -20ºC at 10mg/ml (16ul BSA + 24 ul RIPA)
• Prepare the dilutions of BSA (standard curve):

• Dilute samples 1/10 (in RIPA) and add 5 ul of each sample (or standard) to the wells –> do it for duplicate
• Prepare reagent A: 20 ul reagent S in 1 ml reagent A
• Add 25 ul of reagent A+S in all the wells
• Add 200 ul of reagent B in all the wells
• Mix plate and incubate it for 15 min at RT (coated from the light)
• Read the plate at 650-750 nm
• Calculate the protein concentration