GFP, or green fluorescent protein, is a chemiluminescent protein derived from Aequorea jellyfish that was first discovered by Osamu Shimomura. It was soon after established that the emission spectra of GFP was right around 509nm, or the ultraviolet color range. The GFP gene is often used to form expression constructs in order to closely follow protein behavior, cellular differentiation, protein localization and more. The following articles employed a GFP antibody in conjunction with various other GFP construct techniques to strengthen their research findings.
Immunocytochemistry/Immunofluorescence: GFP Antibody [NB600-308] - Analysis of GFP in transgenic mouse pancreas in OCT. Verified customer review from 1DegreeBio.
Zeng et al used a GFP antibody in transgenic C. elegans samples in their research of the protective effects of xyloketal-derived small molecules in Huntington’s disease (HD). First, Zeng et al generated a polyQ fused to GFP C. elegans model in muscle cells. The polyQ to GFP fusion was decided upon due to the elongated polyQ encoding CAG repeat in the IT15 gene that has been established with Huntington’s disease. In addition, xyloketal B has demonstrated neuroprotection in a variety of neurological disorders. After it was established that the polyQ-GFP construct auto fluoresced under a fluorescent microscope, the protective effects of xyloketal’s were investigated using a GFP antibody. First, the C. elegans were analyzed via fluorescent microscopy after treatment of a number of protective compounds and did show that green polyQ foci were reduced. In order to strengthen this finding, a GFP antibody was used on these same treated and control lysate samples in order to perform densitometry.
Next, Liu et al used a GFP antibody in order to better understand the role of coiled-coil protein Rng10 in septum formation during yeast cytokinesis. The GFP antibody was used in a variety of experimental methods. For starters, cell lysis was observed in a haploid strain of Rng10 depleted yeast. In order to determine whether the lysis was a result of defects in the P.M., fluorescence loss in photobleaching, or FLIP, was utilized. Additionally, a GFP antibody was used in immunoprecipitation assay followed by western blot. First, the mECitrine-tagged Rga7 protein complex was pulled down using GFP coated Dynabeads. Next, the samples were separated and tested with the GFP antibody in western blot.
Another use of the GFP antibody came from Yang et al in their research of how the transplantation of embryonic stem cells improves the regeneration of periodontals. Specifically, GFP was utilized by creating a porcine embryonic stem cell (pES) construct with collagen that expressed GFP. After three months of collagen healing to the damaged tissue, the GFP antibody was used in immunohistochemistry (IHC) in order to track the GFP tagged ES cells. While the IHC results showed that ES cells had differentiated into new periodontal ligaments, more research is needed to further elucidate this pathway.
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PMID: 9759496
PMID: 27385337
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