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Calculation worksheets for LanthaScreen® Kinase Activity Assay


3X Tracer (~3X Kd):

Tracer: Stock = nM
1X tracer = nM
3X tracer = nM

To prepare μL ____ :



V1          x         C1        =         V2         x             C2

                      [Initial]                                             [Final 3X Kd]
Tracer
V1          x       ____nM    =      ____ μL     x         ____ nM

V1 = ___ μL


Buffer: _____ μL – _____ μL = _____ μL 1X Kinase Buffer A

Add the calculated volume of tracer to the calculated volume of 1X Kinase Buffer A.

Note:
Tracer Kd values for Invitrogen™ kinases are shown in the detailed protocol


3X Kinase/3X Antibody:

Kinase:   _____  Molecular weight = _____   kDa (from kinase COA) = μg/nmol

_____mg         x   1,000 μ
g    x    1 nmol    =    ______ nmol
            mL                1 mg                ___μg                       mL


____  nmol         x   1,000 ml    x   ___nmol    =    ______ nM
            mL                   1 L                          L                   



Kinase stock = ____ nM
1X = 5 nM
3X = 15 nM

Antibody stock =  ____ μM = ____ nM
1X = 2 nM
3X = 6 nM


Note:
If V1 is less than 2 μL, perform an intermediate 1:10 dilution. We suggest taking 2.0 μL of kinase and adding it to 18 μL of 1X Kinase Buffer A to prepare 20 μL of diluted kinase

Add the calculated volume of tracer to the calculated volume of 1X Kinase Buffer A.

To prepare 1,500 μL:



V1                x               C1              =              V2               x                C2

                                  [Initial]                                                       [Final 3X]
Kinase V1                x           _____nM         =        1,500  μL       x          _____nM

V1  = _____ μL
Antibody V1                x           _____ nM    =            1,500  μL        x         _____μM

V1  = _____ μL


Buffer:  1,500 ____ μL – ____ μL – ____ μL = ____ μL 1X Kinase Buffer A

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LT129       1-Jan-2010