.
Common cycling parameters:
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 0.5-1 minutes
Annealing 45 to 68 °C 0.5-1 minutes
Extension 72 °C 1-2 minutes
.
Common cycling parameters:
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 0.5-1 minutes
Annealing 45 to 68 °C 0.5-1 minutes
Extension 72 °C 1-2 minutes
for fipronil at 5 ng/mL in the
final extract (Figure 3). Average recovery for spiked
replicates was 94%, with an RSD of 8%, falling well
within the generally acceptable ranges of 70-120%
recovery and
.
Common cycling parameters:
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 0.5-1 minutes
Annealing 45 to 68 °C 0.5-1 minutes
Extension 72 °C 1-2 minutes
.
Common cycling parameters:
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 0.5-1 minutes
Annealing 45 to 68 °C 0.5-1 minutes
Extension 72 °C 1-2 minutes
immunoblotting.
Hypoxia-inducible factor-1 (HIF-1) is a heterodimer
composed of a 120 kDa HIF-1-α subunit complexed
with a 91- to 94 kDa HIF-1-β subunit. The predicted
826-amino acid HIF-1-α contains a bHLH
(1988).
4. Robb, R. J., et al. Proc. Natl. Acad. Sci. USA 81,
6486 (1984).
5. Gillis, S., et al. J. Immunol. 120, 2027 (1978).
6. Fletcher, M., and A. L. Goldstein. Lymphokine Res.
was calculated using the Excel file on the ISO
website (http://standards.iso.org/iso/16140)
100% 94%
IMS method
% of recovery Use 2 different selectivity
agar plates
Direct spread of
the enrichment
intracellular protein aggregation
and cell death by an oxygen-dependent
mechanism. FEBS Lett., 542(1-3), 89-94 (2003).
10. Kimura, T., et al., Quantity and quality control of
gastric proton pump in the endoplasmic
sections).
Hypoxia-inducible factor-1 (HIF-1) is a heterodimer
composed of a 120 kDa HIF -1-α subunit complexed
with a 91- to 94 kDa HIF-1-β subunit. The predicted
826-amino acid HIF -1-α contains a bHLH
1,000 extractions
1,000 amplifications
Extraction
Solution E7526 120 ml 120 ml 120 ml
Dilution Solution D5688 120 ml 120 ml 120 ml
Extract-N-Amp
PCR Ready Mix
or SYBR Green
Extract-N-Amp
PCR
PCR conditions were as follows:
42 °C for 15 minutes; 94 °C for 3 minutes (initial denatu-
ration and JumpStart Taq activation); and 45 cycles of 94 °C
15 seconds and 60 °C 1 minute, with data collection
Common cycling parameters
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 30 seconds
30-35
Annealing 45 to 68 °C 30 seconds
Extension 72 °C
Common cycling parameters
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 30 seconds
30-35
Annealing 45 to 68 °C 30 seconds
Extension 72 °C
1-2
wells), prepare
100 µL of Oxidation Reagent Mix by adding 6 µL of
Chloramine T Concentrate to 94 µL of Oxidation Buffer.
Make a sufficient amount of the Oxidation Reaction Mix
for all of the assay
Common cycling parameters
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 30 seconds
30-35
Annealing 45 to 68 °C 30 seconds
Extension 72 °C
1-2
Common cycling parameters
Step Temperature Time Cycles
Initial
Denaturation 94 °C 3 minutes 1
Denaturation 94 °C 30 seconds
30-35
Annealing 45 to 68 °C 30 seconds
Extension 72 °C