desired buffer. The compound addition is
200 µL/well /96 well plate
7. Run the pH assay by monitoring the fluorescence
at λex = 490/λem = 535 nm (cut off at 515 nm) or
λex = 505/λem = 535 nm
filter that allows passage
of all wavelengths longer than 515 nm will result in a yellow-green emission color, where as a notch filter, (eg. 515-565 nm) will result in a green
emission color. Most filters
volume (in milliliters) of the assay
12.6 = Millimolar extinction coefficient of 4-nitrocatechol at 515 nm.
0.1 = Volume (in milliliter) of enzyme used
and LC FastStart DNA MasterPLUS SYBR Green I* (Cat. No. 03 515 869 001) according
to the procedures within the Instructions for Use.
Fig. 2: Linear control reaction:
Control reaction with PBGD primers
myeloid and
plasmacytoid dendritic cells to lymphoid organs and inflamed skin. J. Exp. Med. 201:509-
515.
Wittamer et al. (2003) Specific recruitment of antigen-presenting cells by chemerin, a novel
45 min,
+37°C
6 Washing: Wash cells twice in PBS 2× 2 min,
+15 to +25°C
7 Analysis: Analyze on a flow cytometer (488nm
excitation using a 515 nm bandpassfilter for
detection) or counterstain
A. M., Mol. Cell Endocrinol., 49, 1-16,
(1987).
7. de Almeida, et al., J. Cell Sci., 107, 507-515 (1994).
8. Wilkie, T. M., and Yokoyama, K. S., Soc. Gen.
Physiol. Ser., 49, 249-270 (1994).
9
minutes at -20°C.
(iii). Wash the cells with PBS, centrifuging at 200 x g for IO minutes.
Fixation method B
(i). Incubate l-2 x 1 06 cells in 100% methanol for 30 minutes at -20°C.
(ii). Wash the
optimal working dilutions
by titration test.
References
1. Mori, Y. et al., Neuroreport, 9, 507-515 (1998).
2. Wes, P.D. et al., Proc. Natl. Acad. Sci. USA, 92,
9652-9656 (1995).
3. Harteneck, C., et
A. M., Mol. Cell Endocrinol., 49, 1-16,
(1987).
7. de Almeida et al., J. Cell Sci., 107, 507-515 (1994).
8. Wilkie, T. M., and Yokoyama, K. S., Soc. Gen.
Physiol. Ser., 49, 249-270 (1994).
, S., Neuroscience (1992) 47:873-896.
Sherry, D. and Ulshafer, R., Visual Neuroscience (1992) 8:515-529.
Sherry, D. and Ulshafer, R., Visual Neuroscience (1992) 9:313-323.
Kalloniatis, M. and Fletcher
Detection Kit
LightTyper 384 Instrument 1 instrument 03 357 414 001
MagNA Pure LC DNA 1 kit 03 310 515 001
Isolation Kit – Large Volume
MagNA Pure LC DNA 1 kit 03 186 229 001
Isolation Kit II (Tissue)
MagNA
physicochemical data for anti-dopamine and anti-p-tyramine antibodies. Molecular Immunology,
(1984) 21, 515-522.
2) Mc RAE-DEGUEURCE A. and GEFFARD M. One perfusion mixture for immunocytochemical detection