The green fluorescent protein (GFP), originally discovered in the jellyfish, Aequorea victoria, is composed of 238 amino acids and has a molecular weight of 27kD. GFP is in the shape of a cylinder, comprising 11 strands of beta-sheet outside, a single alpha helical strand containing the chromophore running through the center, and short helical segments on the ends. GFP transduces the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. GFP has been widely used as a tracer in various diciplines of biology research, such as fluorescence microscopy, reporter gene expression, and in vivo imaging. Laboratory mutation of GFP generated the widely used analog GFPuv, which has brighter fluorescence and more desirable expression properties.
Applications:
Suitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution:
Direct ELISA: 0.5-1ug/ml Detection limit for rGFPuv ~ 1ng/well.
Western Blot: 1-2ug/ml Detection limit for rGFPuv ~ 5ng/lane under non-reducing and reducing conditions.
Immunoprecipitation: 1ug/ml
Optimal dilutions to be determined by the researcher.
Storage and Stability:
Lyophilized powder may be stored at -20 degrees C. Reconstitute to nominal volume by adding sterile 40-50% glycerol, PBS. Aliquot and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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