Littelfuse Inc.
L'immagine può essere rappresentativa.
Vedi le specifiche per i dettagli del prodotto.
GFP
vedi dettagli Schede tecniche

GFP (Green Fluorescent Protein)

Product Overview

GFP belongs to the category of fluorescent proteins and is widely used in molecular biology, cell biology, and biochemistry research. Its main characteristics include its ability to fluoresce green when exposed to blue or ultraviolet light. GFP is typically packaged as a purified protein or as a plasmid DNA encoding the protein. The essence of GFP lies in its unique ability to label and track specific proteins or cells within living organisms. It is available in various packaging options and quantities to suit different research needs.

Specifications

  • Excitation wavelength: 395 nm
  • Emission wavelength: 509 nm
  • Molecular weight: 27 kDa
  • Purity: >95%
  • Storage: -20°C

Detailed Pin Configuration

GFP consists of 238 amino acids and forms a beta-barrel structure with an internal chromophore that emits green fluorescence when excited by light.

Functional Features

  • Labeling and tracking of proteins and cells in live samples
  • Non-toxic to cells and organisms
  • Stable under physiological conditions
  • Compatible with various imaging techniques

Advantages and Disadvantages

Advantages

  • Versatile tool for visualizing biological processes
  • Non-invasive labeling of live cells
  • High photostability

Disadvantages

  • Relatively large size may interfere with protein function
  • Limited brightness compared to some synthetic dyes

Working Principles

GFP's working principle is based on the autocatalytic formation of its chromophore within the beta-barrel structure. When exposed to blue or ultraviolet light, the chromophore emits green fluorescence, allowing for visualization and tracking of the labeled proteins or cells.

Detailed Application Field Plans

GFP finds extensive applications in various fields, including: 1. Cell Biology: Visualizing organelles, protein localization, and cell migration. 2. Molecular Biology: Monitoring gene expression, protein-protein interactions, and protein turnover. 3. Neuroscience: Labeling neurons and studying synaptic activity. 4. Biochemistry: Analyzing protein folding and dynamics.

Detailed and Complete Alternative Models

Several alternative models to GFP are available, each with its own unique features and applications. Some popular alternatives include mCherry, YFP (Yellow Fluorescent Protein), and RFP (Red Fluorescent Protein). These alternatives offer different excitation and emission spectra, allowing for multiplex imaging and expanded experimental possibilities.

In conclusion, GFP is a valuable tool in biological research, offering non-invasive and versatile labeling capabilities. While it has certain limitations, its widespread use and impact in various scientific disciplines make it an indispensable component of modern research methodologies.

[Word count: 366]

10 domande e risposte comuni relative all'applicazione di GFP nelle soluzioni tecniche

  1. What is GFP?

    • GFP stands for Green Fluorescent Protein, a protein derived from jellyfish that emits green light when exposed to blue or ultraviolet light.

  2. How is GFP used in technical solutions?

    • GFP is commonly used as a fluorescent marker in biological research and biotechnology applications to track gene expression, protein localization, and cell behavior.

  3. What are the advantages of using GFP in technical solutions?

    • GFP allows non-invasive visualization of cellular processes, enabling real-time monitoring without disrupting normal cellular function.

  4. Can GFP be used in live cell imaging?

    • Yes, GFP is frequently used in live cell imaging to study dynamic processes within living cells, tissues, and organisms.

  5. Are there different variants of GFP available for specific applications?

    • Yes, there are numerous GFP variants with altered spectral properties, stability, and localization signals to suit specific experimental needs.

  6. What are the challenges associated with using GFP in technical solutions?

    • Challenges include potential cytotoxicity, photobleaching, and interference with cellular processes due to overexpression of GFP.

  7. How can GFP be incorporated into fusion proteins for visualization?

    • GFP can be genetically fused to a protein of interest, allowing researchers to visualize the localization and dynamics of the fused protein within cells.

  8. Can GFP be used for protein-protein interaction studies?

    • Yes, GFP-based assays such as bimolecular fluorescence complementation (BiFC) can be employed to study protein-protein interactions in living cells.

  9. What techniques are commonly used to detect GFP fluorescence?

    • Techniques such as fluorescence microscopy, flow cytometry, and fluorescence spectroscopy are often used to detect and quantify GFP fluorescence.

  10. Are there any alternative fluorescent proteins that can be used alongside or instead of GFP?

    • Yes, other fluorescent proteins such as mCherry, TagRFP, and mTurquoise are commonly used alongside or instead of GFP, offering different spectral properties and applications.