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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 μg 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