C.B.S. Scientific HTLE-7002 User Manual
Page 17
C. B. S. Scientific 17
HTLE-7002
7.
To apply the samples and dye, place a
marked TLC plate on a flat surface.
Remove 1-2
μl of dye mix using the
Gilson micropipetor and “spot” a single
drop (0.3-0.5
μl) on the mark for the dye
(Fig. F).
8.
Dry the spot using an airstream from a filtered air line fitted with a glass pipet at the end to focus the flow. Make sure
never to touch the plate with either the air nozzle or pipet tip, as glass will scratch the cellulose. Before applying the
sample centrifuge it for 2min to pellet any particulate material. Sample volumes are minimally 5
μl range. Usually the
number of cpm needed will determine the amount of each sample loaded. For either C
14
or S
35
one would like to load at
least 1,000cpm and possibly up to 20,000cpm depending on the number of labeled peptides. For P
32
you should try to
load at least 100-200cpm and up to 1,000cpm for proteins that are multiply phosphorylated .
9.
Use the pipet to take up the determined sample volume to be spotted, making sure not to pick up any debris from the
bottom of the tube.
10.
Spot single 0.3-0.5
μl drop on the sample
mark. ( Fig.G)