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Protein Expression & Imaging
The World of Fluorescent Proteins
A comprehensive guide to fluorescent markers, their spectral properties, and the highly specific antibodies used to detect them across applications.
Fluorescent proteins have revolutionized molecular and cellular biology, providing incredibly powerful tools for visualizing and understanding dynamic biological processes in real time.
Derived originally from naturally occurring proteins found in marine organisms, these fluorescent markers allow scientists to tag and track proteins, cells, and entire tissue lineages with remarkable precision. The discovery of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria paved the way for the development of a vast array of colored fluorescent proteins through advanced genetic engineering.
While intrinsic fluorescence is useful for live imaging, highly specific antibodies against these proteins enable researchers to precisely detect, quantify, and analyze these markers in fixed tissues, cell lysates, and flow cytometry.
1. Fluorescent Protein Spectral Guide
Understanding the excitation and emission spectra of your chosen tag is critical for avoiding spectral overlap in multi-color experiments. Below is a comprehensive reference guide linking to our specific detection reagents.
| Protein Name | Excitation (nm) | Emission (nm) | Visual Color |
|---|---|---|---|
| EBFP | 380 | 440 | |
| ECFP | 433 | 475 | |
| EGFP | 488 | 509 | |
| mBanana | 540 | 553 | |
| mOrange | 548 | 562 | |
| tdTomato | 554 | 581 | |
| mTangerine | 568 | 585 | |
| mStrawberry | 574 | 586 | |
| mCherry | 587 | 610 | |
| mGrape1 | 595 | 620 | |
| mRaspberry | 596 | 625 | |
| mPlum | 590 | 648 |
2. The Foundational Fluorescent Proteins
GFP, YFP, and RFP are the cornerstone tools in fluorescent imaging. These proteins remain fundamental for visualizing and studying cellular and molecular processes globally. While intrinsic fluorescence is useful for live-cell tracking, researchers universally rely on specific antibodies against these proteins for comprehensive downstream assays.
GFP (Green Fluorescent Protein)
GFP natively fluoresces green and has been extensively modified to improve its brightness and stability, resulting in variants like eGFP. Anti-GFP antibodies are perhaps the most heavily utilized tag antibodies in modern biology.
YFP (Yellow Fluorescent Protein)
A derivative of GFP, YFP emits yellow fluorescence and is particularly useful in multi-color labeling experiments and highly sensitive FRET (Förster Resonance Energy Transfer) assays.
RFP (Red Fluorescent Protein)
With its far-red emission, RFP is ideal for deep-tissue imaging, minimizing background autofluorescence, and reducing cellular phototoxicity.
3. tdTomato: The Bright Red Beacon
tdTomato is a standout red fluorescent protein known for its unparalleled brightness and photostability. Derived from a mutant of the Discosoma sp. red fluorescent protein (DsRed), tdTomato matures incredibly quickly, making it ideal for live-cell imaging and long-term developmental studies.
The tandem dimer contains two copies of dTomato linked by a 12-amino acid linker. Due to its pair of chromophores, tdTomato is extremely bright and exhibits unique light stability. The biggest disadvantage of tdTomato is that it is a large molecule and may interfere with the folding of fusion proteins in some cases.
Primary Antibody Applications
Researchers frequently use Biorbyt’s tdTomato Antibody (orb182397) for comprehensive downstream validation:
- Western Blotting: Detects tdTomato-tagged proteins, definitively verifying their expression levels in cell or tissue lysates.
- Immunohistochemistry (IHC): Visualizes expression in fixed tissues or transgenic mouse models (where native fluorescence is lost during fixation), providing spatial context.
- Immunoprecipitation (IP): Enriches and isolates tdTomato-tagged proteins from complex mixtures to study protein-protein interactions.
4. mCherry & mOrange Spotlights
mCherry: The Versatile Red Marker
mCherry is another incredibly popular red fluorescent protein derived from DsRed. It is strictly monomeric, making it highly suitable for direct fusion with other proteins without disrupting their native structural function. Its robustness and versatility have made it a go-to marker for studying protein-protein interactions and monitoring cellular events.
View highly validated anti-mCherry Antibody →mOrange: The Bright Orange Option
mOrange offers bright orange fluorescence, making its distinct color profile highly valuable for multi-color labeling experiments. This allows researchers to visualize multiple cellular components simultaneously without spectral bleed-through. mOrange's brightness and stability have made it a favorite in studies requiring precise and clear imaging.
View anti-mOrange Monoclonal Antibody →5. mScarlet & Other Engineered Variants
mScarlet is a newer addition to the family of red fluorescent proteins, engineered for exceptional brightness and maturation speed. Its high quantum yield and incredible photostability make it an excellent choice for super-resolution microscopy and other advanced imaging techniques. We provide robust mScarlet antibodies to confirm expression and conduct Co-IP interaction studies.
Browse Other Specialized Tags
Biorbyt offers a wide range of high-quality antibodies targeting both classical and highly specialized fluorescent variants.
Fluorescent Marker
CFP Antibody
Detect Cyan Fluorescent Protein expression reliably across Western Blot and IF applications.
Fluorescent Marker
Venus Antibody
Highly specific antibody against Venus, an exceptionally bright variant of YFP used heavily in FRET.
Fluorescent Marker
mRuby Antibody
Validate the expression of mRuby, a bright, monomeric red fluorescent protein.
Fluorescent Marker
mStrawberry Antibody
Monoclonal antibody targeting the mStrawberry variant for robust protein detection.
Fluorescent Marker
mPlum Antibody
Detect mPlum, a far-red fluorescent protein highly suited for deep tissue imaging.
Fluorescent Marker
mBanana Antibody
Specifically target and validate the expression of the mBanana fluorescent variant.