record the increase in A340nm for approximately 5 minutes.
2EWDLQ�WKH� $340nm/minute using the maximum linear rate for both the Test and Blank.
05/10/94 Page 3 of 4
SPSURC13
Enzymatic Assay of
stand out, to be prominent).
{http://www.pnas.org/cgi/reprint/94/23/12425.pdf}.
Western blotting: Recommended primary antibody dilution: 1-5 µg/ml in 0.3% Tween in
PBS. Sample preparation: Standard Laemmli
primers is: first
cycle: 5 min at +94°C; 2 min at +50°C; 3 min at +72°C; subsequent
cycles (up to 40 cycles): 1 min at +94°C; 2 min at +50°C; 3 min at
+72°C; last cycle: 1 min at +94°C; 2 min at
Total hardness 120 µS/cm.
LSI calculation sheet
(example)
Test result
This test is based on ASTM standard calculation procedure (D3739-94).
Feed water Value Unit
parameters
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
is
required for each reaction well. For each well, add
6 µL of Chloramine T Concentrate to 94 µL of
Oxidation Buffer and mix well.
Diluted DMAB Reagent – 100 µL
.
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
.
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
.
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.