As noted above, many of the GFP variants utilize filters that are provided standard with the FL600.
If wt GFP is being measured, it is recommended that the 400/20 excitation filter be used in conjunction with a 508/20 emission filter, both of which are custom filters stocked by Bio Tek Instruments.
Here we characterize the excitation and emission spectra of some of the commonly used variants of GFP and recommend appropriate filter sets for detection and quantitation using the FL600 microplate fluorescence reader.
In terms of filter sets for the FL600 microplate fluorescence reader, the most commonly used mutations to the wt GFP protein can be categorized into three groups: (1) the red-shifted variants; (2) the wild type like variants; and (3) the blue emitting variants.
With her is her latest date, Zack, a less than bright guy.
This encounter with Leonard and Zack together demonstrates to Penny that she can't go back to dating the type of guys she used to date: good looking but dumb.
I got nowhere else to be", at which point Bernadette tells Sheldon to just come in. Bernadette: Well, we're really happy for you and we know how much- Amy: I do know what to say.
Greeting the ladies, Sheldon tells them " Amy: Okay, what is going on? Penny: Yes, he said he wants to do something to show you how much you mean to him. In a drunken stupor later, Penny takes some action to reclaim what she's missed no longer dating Leonard. Meanwhile, Howard and Raj believe the answer to Leonard's Penny conundrum is to find another woman for him.After Leonard rejects Howard's idea of using on-line matchmaking services, Raj thinks an interesting experiment would be to see if such a service would produce anyone for Sheldon.Note that although the peak for excitation of wt GFP, GFPuv, and the Stemmer mutant is closer to the 400/30 excitation filter, the 360/40 filter that is a standard filter for the FL600 provides quite adequate fluorescent signal.The red-shifted mutants have been developed for use with the standard "fluorescein" filter set and as such utilize the 485/20 excitation and 530/25 emission filter set.Using the "fluorescein" filter set (485/20 excitation, 530/25 emission) when quantitating wt GFP results in a 20% loss of signal, when compared to the recommended 400/30 excitation, 508/20 emission filter set (Figure 2B).Tags: Adult Dating, affair dating, sex dating