to give a cell density of 0.9 to 1.1
x 106 viable cells/mL
10. Place the flask in the incubator. CO2 and humidity
are not required. Set the temperature to 27 °C. Set
agitation to 130–140 rpm on a
cell growth rates (doubling times of 20-24 hours), high
cell densities of >20 x 106 cells/mL for Sf21 and >10 x
106 cells/mL for Sf9, while maintaining high cell viability
and high recombinant protein
Agar mod.
Yeast
per ml 102 103 104 105 106
Fungi
+ ++ +++
slight moderate heavy
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buffer solution.
27. Using citrate buffer solution, adjust the cell concentration to 2.5 x 106 cells/ml.
Staining Procedure
1. Place 200 l of the prepared cell suspension (2.5 x 106 cells/ml) into
isolation of HSC.
27. Conduct a viable cell count. As a guide, 2 femurs, 2 tibias, and 2 iliac crests from a single 6-8
week old male C57Bl/6J mouse should yield 80-100 x 106 viable nucleated cells
Vaccines Against Cancer. Nat Rev
Immunol, 5(4), 296-306 (2005)
2. Lanzavecchia, A. and Sallusto, F., Regulation of T
Cell Immunity by Dendritic Cells. Cell, 106(3), 263-
266 (2001)
3. Mellman, I. and
cross reactivity with
human immunoglobulins.
APPLICATIONS:
Immunofluorescence: ≤ 1 µg/106 cells
ELISA
Optimal working dilutions must be determined by the end user.
SPECIES REACTIVITY
kinase type IV. Genomics, 4, 21-
27 (1989).
2. Kang, H. et al., An important role of neural activity-
dependent CaMKIV signaling in the consolidation of
long-term memory. Cell, 106, 771-783 (2001).
BKR
methods and characterization of
viral aerosol". University of Florida, Ph.D.
dissertation, pp. 27, 106 (2012).
14. Robinson, Camilla, "Utilisation of mucin sulphur
by Pseudomonas aeruginosa -
with PBS, being careful not
to dislodge any of the cells.
3. Discard the PBS.
4. Add cell lysis buffer (106 – 107 cells/mL).
For cells in suspension
1.
stimulation.
2. Count cells and place 5X 106 cells into a sample tube.
3. Spin down cells at 2500 rpm (600Xg) for 3 minutes and discard buffer.
4. Resuspend the cells in 1 mL of
stimulation.
2. Count cells and place 5X 106 cells into a sample tube.
3. Spin down cells at 2500 rpm (600Xg) for 3 minutes and discard buffer.
4. Resuspend the cells in 1 mL of