GFP Protein, Aequorea victoria (His)

The green fluorescent protein (GFP), Aequorea victoria is a protein that exhibits green fluorescence when exposed to light in the blue to ultraviolet range, found in organisms including Aequorea victoria, corals, sea anemones, zoanithids, copepods and lancelets. GFP Protein has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm, while its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. GFP Protein acts as an energy transfer receptor and actively transduces the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer, which is used as expression of reporter genes. GFP Protein, Aequorea victoria (His) is a recombinant jellyfish Aequorea victoria GFP protein with N-6*His labeled tag that consists of 238 amino acids, which is expressed in E. coli ^{[1][2][3][4][5][6][7]}.

GFP fluorescence occurs when aequorin interacts with Ca²⁺ ions, inducing a blue glow in Aequorea victoria. Some of this luminescent energy is transferred to the GFP, shifting the overall color towards green, making GFP protein enabling the visualization of specific proteins and cellular structures in living organisms^[3].
GFP Protein has a β barrel structure consisting of eleven β-strands with a pleated sheet arrangement, with an α helix containing the covalently bonded chromophore 4-(p-hydroxybenzylidene)imidazolidin-5-one (HBI) running through the center^[4].
In a live cell imaging experiment, on the one hand, GFP Protein directly attaches to a protein of interest to indicate a successful transfection of a gene of interest. On the other hand, GFP Protein that containing a mutation where the fluorescence will change from green to yellow over time is used to study the state of protein production such as recently activated, continuously activated or recently deactivated^[6].
GFP Protein is used widely in cancer research to label and track cancer cells. GFP Protein-labelled cancer cells have been used to model metastasis, the process by which cancer cells spread to distant organs^[7].

GFP Protein (Aequorea victoria) acts as a pollutant marker. There is a decrease in both GFP Protein and cellular density as pollutant levels increases, indicating that cellular activity has decreased^[5].

Zebrafish that are injected with GFP Protein (Aequorea victoria) are approximately twenty times more susceptible to recognize cellular stresses than zebrafish that are not injected with GFP Protein (Aequorea victoria)^[5].
Tumor dissemination and metastasis occur at distinct times during tumor progression in the bi-transgenic MMTV-PyMT and β-actin-enhanced green fluorescent protein (GFP) mice^[7].

Prepare a 150 μL recombinant GFP with various concentrations (0.00074 nM - 6.21 μM) in assay buffer and equilibrate to 25°C. Read at excitation wavelengths 485 nm and emission 535 nm.

Others

E. coli

N-6*His

P42212 (M1-K238)

/

MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK

Approximately 27-31 kDa

• 
  Greater than 90% as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized a 0.2 μm filtered solution of 20 mM Tris-HCl, 0.5 M NaCl, 6% Trehalose, pH 8.0 or 50 mM Tris, 300 mM NaCl, 5% trehalose, 5% mannitol and 0.01% Tween 80, pH 7.4 or PBS, pH 7.4, 8% trehalose.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

Stored at -20°C for 1 year, protect from light. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.

• [1]. Prendergast FG, et al. Chemical and physical properties of aequorin and the green fluorescent protein isolated from Aequorea forskålea. Biochemistry. 1978 Aug 22;17(17):3448-53.  [Content Brief]

  [2]. Phillips GJ. Green fluorescent protein-a bright idea for the study of bacterial protein localization. FEMS Microbiol Lett. 2001 Oct 16;204(1):9-18.  [Content Brief]

  [3]. Morise H, et al. Intermolecular energy transfer in the bioluminescent system of Aequorea. Biochemistry. 1974 Jun 4;13(12):2656-62.  [Content Brief]

  [4]. Ormö M, et al. Crystal structure of the Aequorea victoria green fluorescent protein. Science. 1996 Sep 6;273(5280):1392-5.  [Content Brief]

  [5]. Song Y H, et al. Noninvasive monitoring of environmental toxicity through green fluorescent protein expressing Escherichia coli[J]. Korean Journal of Chemical Engineering, 2016, 33: 1331-1336.

  [6]. Terskikh A, et al. "Fluorescent timer": protein that changes color with time. Science. 2000 Nov 24;290(5496):1585-8.  [Content Brief]

  [7]. Kouros-Mehr H, et al. GATA-3 links tumor differentiation and dissemination in a luminal breast cancer model. Cancer Cell. 2008 Feb;13(2):141-52.  [Content Brief]