Re: [DIYbio] GFP visibility vs concentration?

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IIRC, protein absorption is measured at 280nm? Make sure that's not in a
significant absorption band for your GFP fluorophore, or the fluorophore
might throw off your readings.

That is, I don't think the 280 readings generally assume that the
protein content of the solution has an unusually high absorbance at that
wavelength, but a GFP variant might! :)

On 17/01/14 03:35, kenny kostenbader wrote:
> Nathan, remind me to come back to you when I have a scientific
> heavy-lifting problem on my hands :) that's quite the solution!
>
> Avery, I'd be able to get a value for mg/ml of protein from an absorbance
> reading.
>
> I was hoping to find a set of pictures that compare jars of GFP with
> differing concentrations, something like that. When I get to my own
> samples, I'd be happy to perform some dilutions and post pictures
> somewhere. Someone out there will find that interesting :)
>
>
> Kenny
>
>
> On Thursday, January 16, 2014 7:50:08 PM UTC-5, Avery wrote:
>>
>> or you could get some GFP and serially dilute it until you cant see it.
>>
>> Filters help block the light that causes the excitement, but also make the
>> overall image dimmer.
>>
>> How do you measure GFPs/volume?
>>
>> --A
>>
>>
>> On Thu, Jan 16, 2014 at 6:17 PM, Nathan McCorkle <nmz...@gmail.com<javascript:>
>>> wrote:
>>
>>> you might try finding the dosage (watts of light per area... i.e.
>>> w/cm^2) that photobleaching of GFP occurs at, then from there
>>> determine the maximum fluorescence (watts of light per area, or # of
>>> photons), then find the minimum detection limit for the eye for that
>>> wavelength (watts/area or photons).
>>>
>>> With that you should be able to determine how much light a single GFP
>>> gives off at full brightness, then you can divide the minimum
>>> sensitivity of the eye by that number to get # of GFPs needed. You
>>> could then modify the equations with lower input power, in case you
>>> don't have a super-bright UV light that is capable of photo-bleaching.
>>>
>>> On Thu, Jan 16, 2014 at 2:26 PM, kenny kostenbader
>>> <kenst...@gmail.com <javascript:>> wrote:
>>>> Hi All,
>>>>
>>>> I was wondering how much GFP (in aqueous solution) would be required to
>>> glow
>>>> bright enough to see with the naked eye? Or does it depend more on the
>>>> intensity/color of the excitation lighting?
>>>>
>>>>
>>>> Thanks!
>>>> Kenny
>>>>
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>>
>>
>