GFP, Chicken Polyclonal, Cat# CPCA-GFP-AP

GFP, Chicken Polyclonal, Cat# CPCA-GFP-AP

Transfected HeK293 cells which overexpressed GFP-fusion protein fused to a nuclear localization sequence  were stained with CPCA-GFP. Most Hek293 cells are not transfected so only the nucleus of these cells can be visualized with a blue DNA stain. Cells which are transfected with GFP are bright green. Staining with CPCA-GFP is shown in red. Red antibody staining is only seen in cells which express GFP, as expected, and the superimposition of green and red results in an orange signal.

Blot of HEK293 cells transfected with pFin-EF1-GFP vector from the laboratory of Dr Susan Semple-Rowland, University of Florida. This vector expresses full length GFP and crude homogenate from these cells was probed with our CPCA-GFP antibody. There is a strong clean band at about 27kDa corresponding to GFP.

Product name Anti-GFP
Description Chicken Polyclonal Antibody to GFP
Reference Code CPCA-GFP
HGNC name
RRID# AB_2572314
Molecular weight ~27 kDa
Immunogen Prot-aceGFP recombinant protein
Isotype IgY
Concentration Antibody is provided as affinity purified antibody at 1mg/ml
Species Reactivity NA
Applications Western blot, ICC/IF, IHC
Storage instructions Shipped on ice. Store at 4°C. For long term storage, leave frozen at -20°C. Avoid freeze / thaw cycles.
Suggestions for use Western blot: 1:1,000-5,000
IF/IHC: 1:1,000-5,000

Green Fluorescent Protein (GFP) is a ~27 kDa protein isolated originally from the jellyfish Aequoria victoria. It is a fluorescent protein with an excitation maximum at 395 nm and emission maximum at 509 nm, so that when excited with blue or UV light, it emits green light (1). The fluorescence of GFP requires only the polypeptide chain and molecular oxygen and no other cofactors, so it can be expressed in fluorescent form in essentially any prokaryotic or eukaryotic cell. GFP has been engineered to produce a vast number of variously colored mutants including blue, cyan and yellow protein derivatives, such as BFP, CFP and YFP etc (2-4). GFP and these derivatives are widely used as fluorescent tracers in transfection and transgenic experiments to monitor gene expression and protein localization in Vivo. GFP was the basis of the 2008 Nobel Prize in Chemistry, awarded to Osamu Shimomura, Martin Chalfie and Roger Tsien, specifically “for the discovery and development of the green fluorescent protein, GFP”.

References:

1. Shimomura O, Johnson FH, Saiga Y. Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. Journal of Cellular and Comparative Physiology 3:223–39 (1962).

2. Ormo M, Cubitt AB, Kallio K, Gross LA, Tsien RY, Remington SJ. Crystal structure of the Aequorea victoria green fluorescent protein. Science 273:1392-95 (1996).

3. Heim R, Prasher DC, Tsien RY. Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc. Natl. Acad. Sci. USA 91:12501-04 (1994).

4. Lelimousin M, Noirclerc-Savoye M, Lazareno-Saez C, Paetzold B, Le Vot S, Chazal R et al. Intrinsic dynamics in ECFP and Cerulean control fluorescence quantum yield. Biochemistry 48:10038–10046.

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