You have no items in your shopping cart.
GFP & EGFP Fluorescent Proteins | GFP Antibodies, Recombinant Proteins & Live-Cell Imaging
Green fluorescent protein (GFP) and enhanced green fluorescent protein (EGFP) are among the most widely used fluorescent proteins in molecular and cell biology. They are commonly used for live-cell imaging, protein localization, fusion protein studies, reporter gene assays, transfection monitoring, and stable cell line generation.
Overview
Green fluorescent protein (GFP) and enhanced green fluorescent protein (EGFP) are among the most widely used fluorescent proteins in molecular and cell biology. They are commonly used for live-cell imaging, protein localization, fusion protein studies, reporter gene assays, transfection monitoring, and stable cell line generation.
Although wild-type GFP played a foundational role in fluorescent protein research, EGFP has become the preferred green fluorescent protein variant for most modern mammalian cell experiments. EGFP provides stronger fluorescence, faster maturation, improved folding at 37°C, and more reliable mammalian cell expression.
GFP and EGFP are also widely used in cell tracking, tumor imaging, and tumor microenvironment studies, often in combination with additional cell-type or immune markers.
Biorbyt provides a range of anti-GFP antibodies, anti-EGFP antibodies, GFP nanobody/VHH reagents, and recombinant GFP proteins for applications including WB, IF, ICC, IHC, IP, Co-IP, flow cytometry, assay development, and antibody validation.
What Are GFP and EGFP?
Green fluorescent protein (GFP) is a fluorescent protein originally identified in Aequorea victoria. In modern molecular and cell biology, "GFP" may refer either to the original wild-type GFP protein or more broadly to GFP-derived fluorescent tags used in protein labeling, cell tracking, reporter systems, and fusion protein studies.
Enhanced green fluorescent protein (EGFP) is an engineered GFP variant developed to improve fluorescence performance and expression in experimental systems. EGFP retains the green fluorescence properties of GFP but includes key modifications such as S65T, F64L, and mammalian codon optimization.
These changes improve:
- Fluorescence brightness
- Chromophore maturation
- Protein folding at 37°C
- Expression efficiency in mammalian cells
- Signal reliability in imaging workflows
In practical research use, many constructs, antibodies, and product descriptions use "GFP" as a general term covering GFP, EGFP, and related GFP-tagged fusion proteins. Therefore, when selecting a GFP antibody or GFP detection reagent, researchers should confirm whether the product has been validated for the specific GFP variant used in their experiment.
GFP vs EGFP: Practical Experimental Differences
The main difference between GFP and EGFP is not their general use, but their experimental performance under standard laboratory conditions.
Wild-type GFP is mainly relevant for historical constructs, legacy vectors, or experimental systems where the original GFP sequence is specifically required. By contrast, EGFP is usually preferred for modern mammalian cell experiments because it was engineered for stronger and more reliable fluorescence.
The S65T mutation improves fluorescence excitation and brightness, while the F64L mutation enhances folding efficiency at physiological temperature. Mammalian codon optimization further supports stronger expression in mammalian systems.
For most modern workflows, EGFP is generally preferred for:
- Live-cell imaging
- Reporter gene assays
- Transfection monitoring
- Lentiviral expression systems
- Stable fluorescent cell line generation
- Protein localization studies
From a detection perspective, many anti-GFP antibodies can recognize EGFP because GFP and EGFP share highly similar amino acid sequences. However, antibody compatibility should always be confirmed from the product datasheet, especially for WB, IF, IHC, IP, Co-IP, ICC, and flow cytometry.
Applications of GFP and EGFP
GFP Fusion Proteins for Live-Cell Imaging
GFP and EGFP are widely used to generate fluorescent fusion proteins for monitoring protein localization and cellular dynamics in living cells. By genetically fusing GFP or EGFP to a protein of interest, researchers can directly visualize intracellular distribution, protein trafficking, organelle localization, and cell morphology in real time.
Common applications include:
- Live-cell fluorescence imaging
- Confocal microscopy
- Time-lapse imaging
- Subcellular localization studies
- Protein trafficking analysis
- Cell migration monitoring
- Organelle imaging
Because EGFP provides stronger fluorescence intensity and improved folding at 37°C, it is generally preferred for mammalian live-cell imaging experiments.
In low-expression systems or fixed-cell workflows, endogenous GFP fluorescence may become weak or partially reduced after sample processing. In these situations, anti-GFP antibodies are often used to improve signal detection and imaging sensitivity.
Reporter Gene and Transfection Monitoring
EGFP is one of the most commonly used fluorescent reporter genes in molecular biology. GFP or EGFP expression allows rapid identification of successfully transfected or transduced cells without additional staining procedures.
Typical applications include:
- Plasmid transfection monitoring
- Lentiviral expression systems
- CRISPR workflow validation
- Stable fluorescent cell line generation
- Promoter activity analysis
- Reporter gene assays
Because EGFP matures efficiently and produces strong fluorescence in mammalian cells, it has become a standard reporter protein in many expression vectors and cell-based assays.
Although GFP fluorescence can often be directly observed, antibody-based detection is commonly used for Western blot validation, fixed-cell immunofluorescence, and low-abundance reporter systems where fluorescence intensity alone may not be sufficient.
GFP in Cell Tracking and Tumor Research
GFP and EGFP are also widely used in cell tracking experiments, including tumor biology, immune cell migration, and tumor microenvironment studies. GFP-labeled cells allow researchers to monitor cell behavior over time and visualize how cells move, expand, or respond to experimental conditions.
Common cell tracking applications include:
- Tumor growth and metastasis monitoring
- Cell migration studies
- Drug response assays
- Tumor-host interaction studies
- Immune cell trafficking
- Tumor microenvironment analysis
In some workflows, GFP reporter systems are combined with additional cell-type markers, such as macrophage or immune cell markers, to study cell distribution and tissue-level interactions.
GFP Antibodies, Nanobodies and Recombinant Proteins
Anti-GFP and anti-EGFP antibodies are widely used to detect GFP-tagged proteins in WB, IF, ICC, IHC, IP, Co-IP, and flow cytometry. Although GFP fluorescence can often be observed directly, antibody-based detection provides higher sensitivity, signal amplification, and confirmation of full-length fusion protein expression.
This is especially useful for:
- Low-expression GFP fusion proteins
- Fixed-cell imaging
- Western blot validation
- Protein pull-down assays
- Reporter gene verification
- GFP-tagged protein detection
Because GFP-family proteins share highly conserved sequences, many anti-GFP antibodies may also recognize EGFP, EYFP, CFP, and related variants. Cross-reactivity depends on the antibody clone, epitope, and application, so researchers should check product validation data before use.
Biorbyt provides a range of GFP and EGFP antibodies, GFP VHH/nanobody reagents, recombinant GFP antibodies, and recombinant GFP proteins for detection, validation, and protein capture workflows.
Recommended GFP & EGFP Antibodies and Proteins
Product Name | SKU | Applications | Size / Format | Price |
|---|---|---|---|---|
GFP Antibody — Broad GFP/EGFP Detection | ELISA, IF, IHC, WB | 1 mg | $1,120 | |
GFP Rabbit Polyclonal Antibody — Literature Supported | ELISA, IF, IHC, WB | 100 μg | $680 | |
EGFP Rabbit Polyclonal Antibody — Literature Supported | ELISA, WB | 100 μg | $300 | |
GFP-tag Monoclonal Antibody — Literature Supported | IP, WB | 30 μl | $110 | |
GFP VHH Single-Domain Antibody — Alpaca-Derived Binder for IP and Protein Capture | ELISA, IP, WB | 50 μg | $290 | |
GFP Recombinant Rabbit mAb — Broad Recombinant Antibody | ELISA, FC, ICC, IF, IHC-Fr, IHC-P, WB | 100 μl | $490 |
Biorbyt also provides recombinant GFP and EGFP proteins for assay development, positive controls, antibody validation, and fusion protein detection workflows. Recommended options include Recombinant GFP Protein, His (orb1516874), GFP Control Protein (orb345957), and Recombinant Enhanced GFP Protein, C-His (orb2961410).
GFP Nanobodies for Protein Capture
GFP nanobodies, also known as GFP VHH single-domain antibodies, are small antibody-derived binders that recognize GFP-tagged proteins with high affinity. Compared with conventional antibodies, GFP nanobody-based reagents can provide rapid binding, low nonspecific background, and efficient enrichment of GFP-tagged fusion proteins.
They are commonly used in:
- Immunoprecipitation (IP)
- Co-immunoprecipitation (Co-IP)
- Protein pull-down assays
- GFP-tagged protein enrichment
- Protein interaction studies
For researchers studying GFP-tagged fusion proteins or protein interaction networks, GFP nanobody-based reagents provide a useful option for protein capture and enrichment workflows.
Related GFP Family Variants
The GFP family includes multiple engineered fluorescent proteins derived from or related to wild-type GFP. These variants differ in fluorescence color, emission peak, brightness, stability, and oligomeric state, making them useful for live-cell imaging, protein localization, multicolor imaging, and FRET assays.
.jpg)
Protein | Color | Structure | Emission Peak (nm) | Brightness | Stability | Notes |
|---|---|---|---|---|---|---|
EGFP | Green | Weak dimer | ~509 | Moderate to high | Moderate | Standard GFP variant for mammalian imaging and reporter assays |
mEGFP | Green | Monomer | ~509 | Moderate to high | Moderate | Monomeric EGFP variant, preferred for fusion protein localization |
EYFP / Venus | Yellow | Weak dimer / monomer | ~527–528 | High | Moderate | Common FRET acceptor and multicolor imaging marker |
ECFP / mTurquoise | Cyan | Monomer | ~475 | Moderate to high | Moderate | Common FRET donor for protein interaction studies |
EBFP / Azurite | Blue | Weak dimer / monomer | ~446–460 | Low to moderate | Moderate | Blue-channel variants for additional imaging channels |
mNeonGreen | Green | Monomer | ~517 | Very high | Moderate to high | Very bright green fluorescent protein from a non-Aequorea source |
mStayGold | Green | Monomer | ~510 | Very high | Very high | Highly photostable green fluorescent protein for long-term imaging |
When choosing a GFP-family variant, researchers should consider color, brightness, photostability, and whether the protein is monomeric, especially for fusion protein studies. Monomeric variants are generally preferred because they are less likely to interfere with protein localization or interaction analysis.
For dual-color or multicolor imaging, GFP-family proteins are frequently combined with red fluorescent proteins such as mCherry and tdTomato, which provide well-separated fluorescence channels for co-localization, cell tracking, and protein interaction studies.
FAQ
What is the difference between GFP and EGFP?
EGFP is an engineered version of wild-type GFP designed to provide stronger fluorescence, faster maturation, improved folding at 37°C, and more efficient mammalian expression. In modern mammalian cell experiments, EGFP is generally preferred for live-cell imaging, reporter assays, transfection monitoring, and stable cell line generation.
Why is EGFP brighter than wild-type GFP?
EGFP contains key improvements such as S65T, F64L, and mammalian codon optimization. These modifications improve fluorescence excitation, chromophore maturation, protein folding, and expression efficiency, making EGFP easier to detect under standard mammalian cell culture conditions.
Why is EGFP commonly used in live-cell imaging?
EGFP provides strong green fluorescence, efficient maturation, and reliable expression in mammalian cells. These properties make it suitable for monitoring protein localization, intracellular trafficking, cell morphology, reporter activity, and long-term live-cell imaging workflows.
Can anti-GFP antibodies detect EGFP?
Yes. Many anti-GFP antibodies can detect EGFP because GFP and EGFP share highly similar amino acid sequences. However, antibody compatibility should always be confirmed from the datasheet, especially for applications such as WB, IF, IHC, ICC, IP, Co-IP, and flow cytometry.
Can GFP antibodies cross-react with YFP or CFP?
Yes. Some anti-GFP antibodies may recognize EYFP, CFP, and related GFP-family variants because these proteins share conserved sequence regions with GFP and EGFP. Cross-reactivity depends on the antibody clone, epitope, and application, so researchers should check product validation data before use.
What is a GFP nanobody?
A GFP nanobody is a small single-domain antibody, often based on VHH technology, that binds GFP-tagged proteins with high affinity. GFP nanobody-based reagents are commonly used for IP, Co-IP, pull-down, and protein capture workflows.
What is the difference between EGFP and mNeonGreen?
mNeonGreen is generally brighter than EGFP and can be useful for high-sensitivity imaging. However, EGFP remains more widely used because it has extensive validation history, broad vector compatibility, and strong antibody support across standard GFP-based workflows.
References
- Tsien, R. Y. (1998). The green fluorescent protein. Annual Review of Biochemistry, 67, 509–544.
- Heim, R., Cubitt, A. B., & Tsien, R. Y. (1995). Improved green fluorescence. Nature, 373, 663–664.
- Cormack, B. P., Valdivia, R. H., & Falkow, S. (1996). FACS-optimized mutants of the green fluorescent protein (GFP). Gene, 173(1 Spec No), 33–38.
- Shaner, N. C., Steinbach, P. A., & Tsien, R. Y. (2005). A guide to choosing fluorescent proteins. Nature Methods, 2(12), 905–909.
- Chudakov, D. M., Matz, M. V., Lukyanov, S., & Lukyanov, K. A. (2010). Fluorescent proteins and their applications in imaging living cells and tissues. Physiological Reviews, 90(3), 1103–1163.
- Rodriguez, E. A., Campbell, R. E., Lin, J. Y., Lin, M. Z., Miyawaki, A., Palmer, A. E., Shu, X., Zhang, J., & Tsien, R. Y. (2017). The growing and glowing toolbox of fluorescent and photoactive proteins. Trends in Biochemical Sciences, 42(2), 111–129.
- Rothbauer, U., et al. (2008). A versatile nanotrap for biochemical and functional studies with fluorescent fusion proteins. Molecular & Cellular Proteomics, 7(2), 282–289.
- Shaner, N. C., et al. (2013). A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum. Nature Methods, 10, 407–409.