2-24-89


                        Protein Assay Lowry Method         

REF STOCK CONCENTRATION: approximately 10g/dl since it changes with every 
                         certified lot.
        A) 1 ml of ref stock  in a total volume of 10.0 ml of dH2O.
               (use a volumetric flask)

           1 ml * 10.2 g/dl divided by total volume * 1000 =[] working stock


CURVE: VALUES USING 10.2g/100 ml stock

        S-1   0.1 ml in 10 ml aprox = 10.2 ug/ml
        S-2   0.2 ml in 10 ml aprox = 20.2 ug/ml
        S-3   0.3 ml in 10 ml aprox = 30.6 ug/ml
        S-4   0.4 ml in 10 ml aprox = 40.8 ug/ml
        S-5   0.5 ml in 10 ml aprox = 51.0 ug/ml
        S-6   0.7 ml in 10 ml aprox = 71.4 ug/ml
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        Remember when converting
        100 ug = 1 mg = .001 g
        2000 ug = 20 mg = .020 g
        1 ug = .001 mg= .000001 g

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Stock Solution                          (make a liter so [] is 20g/L)
        A:   2g/100ml Na2CO3 in 0.1N NaOH   (50cc (2N)/1000)   
        B:   1g/100ml  CuSO4 5H2O
        C:   2g/100ml  K-Na-Tartate
        D:   Folin-Ciocelteu's      Phenol Reagent 2N

WORKING REAGENT

       1) For every 100cc of A add 1cc of B and 1cc of C.
       2) make a 1:1 dilution of D with dH2O ( which makes a 1N working 
                reagent D).

METHOD

        1) 100 ul sample, 100 ul standard, 100 ul blank (NaCl)
                Use same volume of std., unknowns and blanks.
        2) 100 ul + 2 ml working reagent, Vortex and time for 10 min.
        3) After 10 min add 100 ul of D working reagent and Vortex while
          timing for at least 30 min.