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[Biosensis] Glowing Proteins – A Guiding Star for Biochemistry

관리자 2021.10.08 10:40 조회 1150 추천 22
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Glowing Proteins – A Guiding Star for Biochemistry


Dear Researcher,

 

In 2008, the Nobel prize in Chemistry was awarded to 3 scientists, Osamu Shimomura, Martin Chalfie and Roger Y. Tsien, “for the discovery and development of the green fluorescent protein or GFP”. The Nobel prize committee described GFP as a ‘guiding star for biochemistry‘, because this protein has become one of the most important tools in bioscience today. GFP is used in many applications such as fusion tagging, transcription reporting, Foerster Resonance Energy Transfer (FRET), biosensors, optogenetics, fluorescence-activated cell sorting (FACS), as well as a general epitope tag for protein purification.


GFP Characteristics

GFP is a 27 kDa monomeric protein, isolated from the jellyfish Aequorea Victoria. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. It has many mutated allelic forms like blue, cyan and yellow and it can be fused to a broad variety of proteins. Tsien's 'A guide to choosing fluorescent proteins' provides useful information on how to choose the best protein for a given experiment.


GFPs main advantage over conventional fluorescent dyes is that it is non-toxic and can be expressed in living cells enabling the study of dynamic, physiological processes. Further, GFP is a small molecule (238 amino acids) and can remain functional even when fused to a target protein, so it allows protein localization to be monitored in real time.


 Antibodies Against GFP

Biosensis offers four high-quality antibodies against GFP, suitable for Western blots and immunofluorescence analysis to amplify GFP signals in tissues of transgenic animals and cell culture experiments. The choice of four host animals (Chicken, Rabbit, Goat and Mouse) offers maximum flexibility for experimental design. 


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Figure 1: Immunofluorescence analysis of transfected HEK293 cells with a GFP fusion protein construct, in green, and stained with chicken antibody to GFP (C-2102-100, 1:1,000, red). Blue: DAPI nuclear stain. The antibody reveals GFP protein expressed only in transfected cells, and as a result these cells appear golden-yellow in color. Top: merged image; Bottom: individual channels.



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Figure 2: Western blot analysis of HEK293 cell lysates using goat antibody to GFP (G-2103-100, 1:1,000, green). [1] protein standard, [2] non-transfected control cells, [3] cells transfected with GFP, [4] cells transfected with mCherry. A strong band at ~27 kDa corresponds to GFP protein detected only in cells transfected with GFP. This antibody does not cross-react with mCherry protein. The blot was simultaneously probed with a chicken antibody to HSP60 (red, lanes 2-4).



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