Posted by
Michelle Peckham on
Jun 28, 2013; 9:03pm
URL: http://confocal-microscopy-list.275.s1.nabble.com/slowing-down-or-immobilizing-somewhat-purified-fluorescent-protein-tp7580579p7580582.html
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Hi again
This is taken from our Methods paper back in 2003 (Methods 29 (2003)
142152)
Control coverslips, which were sparsely decorated
with single GFP fluorophores, were created by incubating
the coverslip with a low concentration of anti-
GFP antibody, blocking with 1 mg/ml bovine serum
albumin, and then incubating with a solution of about
10nM GFP. Coverslips were then viewed using TIRFM.
Individual GFP fluorophores could be identified as
separate objects that exhibited either one- or two-step
photobleaching (Fig. 7). These control slides were important
in setting up the TIRFM apparatus and in establishing
single-fluorophore sensitivity.
Hope that helps. If you want the whole paper - let me know.
Al best
Michelle
On 28/06/2013 21:06, "Jen Jackson" <
[hidden email]> wrote:
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>Thanks for your response. I would prefer the molecules for the have even
>more mobility so to test temporal resolution. But antibodies are a great
>start- can you recommend any protocol for attaching antibodies to a
>coverslip (just use poly-d-lysine, or?).