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DIG Gel Shift Kit, 2nd generation New formulation containing Terminal Transferase, recombinant.
1 kit, Kit for 20 Oligonucleotide 3�-end labeling reactions with DIG-11-ddUTP, 200, binding reactions, chemiluminescent detection reaction for 20 blot
Contents
Labeling Buffer (5x conc.), 80 �l
CoCl2 Solution (25 mM), 80 �l
DIG-ddUTP Solution (1 mM Digoxigenin-11-ddUTP), 20 �l
Recombinant Terminal Transferase (400 U/�l), 20 �l
Binding Buffer (5x conc.), 800 �l
Control Oligonucletide (ds 39mer, unlabeled, 0.1 �g/�l, 3.85 pmol/�l), 40 �l
DIG-labeled Control Oligonucleotide (ds 39mer, 0.4 ng/�l, 15.54 fmol/�l), 140 �l
Control Factor Oct2A (25 - 75 ng/�l), 40 �l
Poly [d(I-C)] (0.1 �g/�l), 200 �l
Poly [d(A-T)] (0.1 �g/�l), 200 �l
Poly L-lysine (0.1 �g/�l in double-distilled water), 200 �l
Loading Buffer without bromophenol blue, 1 ml
Loading Buffer with bromophenol blue, 1 ml
Anti-Digoxigenin-AP (750 U/ml), 40 �l
CSPD (10 mg/ml), 440 �l
Blocking Reagent, 50 g
Application
Use the DIG Gel Shift Kit to prepare nonradioactive, 3�-end-labeled oligonucleotide probes, so they can be used to detect DNA-protein complexes in a "gel mobility shift assay."
Benefits
Convenient, since the technique does not require special equipment or technology
Reproducible, since DIG-labeled probes are stable indefinitely
Safe, because the assay is nonradioactive
Reliable, because the kit provides DIG-labeled control oligonucleotides to establish that the assay is working
Function-tested with the controls provided in the kit (See "Quality" below.)
Sensitive, since the kit can detect as little as 20 fmol of the control oligonucleotide (after it is labeled according to the kit protocol) |