GFP, Rabbit polyclonal, Cat# RPCA-GFP-AP

GFP, Rabbit polyclonal, Cat# RPCA-GFP-AP

Transfected HEK293 cells which overexpress a GFP-fusion protein including a nuclear localization sequence were stained with RPCA-GFP and viewed in the microscope. 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. On staining with RPCA-GFP in red, cells appear orange. The red antibody staining is only seen in cells which express GFP, as expected, and the superimposition of the green and red signals results in the orange color.

Blot of HEK293 cells transfected with pFin-EF1-GFP vector (lane 1) and non-transfected cells (lane 2) was probed with RPCA-GFP at 1:2,000. There is a strong clean band at ~27 kDa corresponding to GFP in GFP-transfected cells, which is not in non-transfected cells.

Product name Anti-GFP
Description Rabbit Polyclonal Antibody to GFP
Reference Code RPCA-GFP-AP
HGNC name NA
RRID# AB_2572317
Molecular weight ~27 kDa
Immunogen Full length recombinant protein based on the Clontech enhanced GFP
Isotype IgG
Concentration Antibody is provided as affinity purified antibody at 1 mg/ml in 50% glycerol/PBS
Species Reactivity NA
Applications Western blot and 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: 1:5,000

Green Fluorescent Protein (GFP) is a 27 kDa protein isolated originally from the jellyfish Aequoria victoria. It has an endogenous fluorochrome activity with excitation maximum at 395 nm and emission maximum at 509 nm, which is similar to fluorescein (1). GFP fluorescence develops as soon as the protein is expressed and requires only molecular oxygen and no other cofactors. As a result GFP 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, BFP, CFP and YFP (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 (2009).

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