Questions tagged [green-fluorescent-protein]

Green fluorescent protein is a protein of the jellyfish Aequorea victoria which emits bright green light when it excited with blue to ultraviolet light. The protein is widely used in cell biology to tag proteins and to follow up their localization in the cell.

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CRISPR/Cas9 edited E.coli on AFM?

I am doing CRISPR/Cas9 experiment on E. coli. I am introducing recombinant plasmid BPK764 (which carries Cas9 + sgRNA designed and added later in that plasmid) into compentent E.coli cells which ...
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1answer
31 views

Fluorescent protein tags and colocalisation

We want study if 2 proteins A and B are co-located, for that we use 2 FTP(Fluorescent tag proteins) for each protein?and after the expirement these 2 FTP are co-located. Does that mean necessarily ...
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Why do I see so many kinetochores?

I am analysing RPE-1 cells from humans and I do not understand why I see so many kinetochores by immunofluorescence (more than 100 in many cells). They are in prometaphase.
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GFP brightness calculation

So this question is part math, part biology, but I'm asking about the biology part here (or maybe the chemistry part). Basically I'm asking about my attempt to calculate the possible brightness per ...
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Clarification of the brightness of bioluminescent proteins

One basic thing I'm not too clear on, which I would like to be clear on, is the brightness per mass unit of GFP or luciferase. I was having trouble finding a resource that would state the brightness ...
2
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1answer
95 views

What does it mean when a protein has a different excitation spectra than wildtype, but the same emission spectra?

We have a GFP mutant that displays a different excitation spectra with emission at 510nm than the WT. However, their emission profiles with excitation at 490nm are the same, and we do not observe the ...
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985 views

Can I image Coomassie and GFP in gels at the same time with a fluorescence scanner?

I'm working with a GFP-tagged protein and am routinely using a fluorescence imager (GE Typhoon) and a standard optical scanner to capture fluorescent and absorption images, respectively, of my SDS-...
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2k views

Can I elute my GFP-tagged protein off anti-GFP antibody using a peptide?

I would like to perform a modified co-immunoprecipitation assay using a GFP-tagged protein. We are going to bind the tagged protein to anti-GFP antibody then bind that to protein A/G beads, however ...