Bertin, J., et al., Proc. Natl. Acad. Sci. U. S. A. 94,
1172-1176 (1997).
13. Goltsev, Y. V., et al., J. Biol. Chem., 272, 19641-
19644 (1997).
14. Shu, H. B., et al., Immunity, 6, 751-763 (1997)
al., Proc. Natl. Acad. Sci. USA. 94,
1172-1176 (1997).
14. Shu, H. B., et al., Immunity, 6, 751-763 (1997)
15. Inohara, N., et al., Proc. Natl. Acad. Sci. USA, 94,
10717-10722 (1997).
16. Srinivasula
www.sigmaaldrich.com
07
21
.1
14
94
45
70
01
18
2. How to Use this Product
y Version: 18 BM Blue POD Substrate, soluble 3,3’-5,5’-Tetramethylbenzidine (TMB)
4 sigma-aldrich.com
2.
Natl. Acad. Sci. U. S. A. 94,
1172-1176 (1997).
12. Irmler, M., et al., Nature, 388,190-195 (1997).
13. Goltsev, Y.V., et al., J. Biol. Chem.,272:
19641-19644 (1997).
14. Shu, H. B., et al., Immunity
(10) (0.05)
CPM counted = Actual count - background on filters
Revised: 12/27/94 Page 4 of 4
Enzymatic Assay of PROTEIN KINASE CATALYTIC SUBUNIT
Sigma Prod. Nos. P-2645 and P-8289
receptors. Cell, 94, 481-490 (1998).
3. Li, H. et al., Cleavage of BID by caspase 8
mediates the mitochondrial damage in the Fas
pathway of apoptosis. Cell, 94, 491-501 (1998).
4. Zha, J. et
receptors. Cell, 94, 481-490 (1998).
3. Li, H. et al., Cleavage of BID by caspase 8
mediates the mitochondrial damage in the Fas
pathway of apoptosis. Cell, 94, 491-501 (1998).
4. Zha, J. et
12. Bertin, J., et al., Proc. Natl. Acad. Sci. USA, 94:
1172-1176 (1997).
13. Goltsev, Y.V., et al., J. Biol. Chem.,272:
19641-19644 (1997).
14. Shu, H. B., et al., Immunity, 6: 751-763 (1997).
at pH 7.6 at
25°C in the presence of 1.3 mM 2,3-diphosphoglycerate.
Revised: 11/28/94 Page 4 of 4
Enzymatic Assay of PHOSPHOGLYCERATE MUTASE
(EC 5.4.2.1)
FINAL ASSAY CONCENTRATIONS:
receptors, Cell, 94, 481-490 (1998).
3. Li, H. et al., Cleavage of BID by caspase 8
mediates the mitochondrial damage in the Fas
pathway of apoptosis. Cell, 94, 491-501 (1998).
4. Zha, J. et al
receptors, Cell, 94, 481-490 (1998).
3. Li, H. et al., Cleavage of BID by caspase 8
mediates the mitochondrial damage in the Fas
pathway of apoptosis. Cell, 94, 491-501 (1998).
4. Zha, J. et al
CV
QC Low 90 10
QC Mid 88 9.4
QC High 94 8.3
QC Endo 95 15
Objectives
Development of two highly sensitive methods for the quantitation of
IL-4 and IL-13 in the human
catalytic subunit target
94 °C 2 min
98 °C 10 s, 68 °C 7 min (30 cycles)
13.5 kb dystrophin target
94 °C 2 min
98 °C 10 s, 68 °C 14 min (35 cycles)
8.9 kb IGFR2 receptor target
94 °C 2
min
Initial denaturation 94°C for 7 min
Standard PCR Profile Setup Temp. Time (s) Cycles
Denaturation 94°C 10
10Annealing ×1) °C 30
Elongation 68°C ×2 )
Denaturation 94°C 10
25
Annealing
primer-removal
reaction product or control
4. Perform qPCR using the following amplification
conditions:
1 cycle
94 °C, 2.5 minutes
40 cycles:
94 °C, 30 seconds
__ °C, 30 seconds*
72 °C, 30 seconds