Introduction
Fluorescent proteins have revolutionized the field of molecular and cellular biology, enabling researchers to track gene expression, protein localization, and intracellular dynamics in real time. TagRFP (red fluorescent protein) is widely used for live-cell imaging due to its brightness and photostability. To enhance its detection and analysis, researchers employ Goat Anti-TagRFP Polyclonal IgG Antibody, a powerful immunological tool used in western blotting, immunofluorescence, immunoprecipitation, and other biochemical assays (nih.gov).
This article provides a comprehensive overview of Goat Anti-TagRFP Polyclonal IgG Antibody, covering its features, applications, and technical aspects, while referencing authoritative .edu and .gov sources to ensure high-quality scientific credibility.
What is TagRFP?
TagRFP is a red fluorescent protein derived from the sea anemone Entacmaea quadricolor and has been optimized for use in mammalian cells. It offers superior photostability and minimal cytotoxicity, making it ideal for long-term live-cell imaging (nih.gov).
Key Properties of TagRFP:
- Excitation wavelength: 555 nm
- Emission wavelength: 584 nm
- High photostability and brightness (ncbionlm.nih.gov)
- Compatible with various imaging and detection techniques (nih.gov)
Goat Anti-TagRFP Polyclonal IgG Antibody: Features and Benefits
This polyclonal antibody is highly specific for TagRFP and provides robust detection capabilities across multiple applications.
1. High Specificity and Affinity
- Recognizes TagRFP and its fusion proteins without cross-reacting with other fluorescent proteins (nih.gov).
- Produced using immunized goats to ensure strong and polyclonal immune response (fda.gov).
2. Versatility Across Applications
- Western Blotting (WB): Detects TagRFP-tagged proteins in cell lysates (ncbi.nlm.nih.gov).
- Immunofluorescence (IF): Enables visualization of intracellular TagRFP fusion proteins (nih.gov).
- Flow Cytometry (FACS): Used in quantifying TagRFP-expressing cells (nih.gov).
- Immunoprecipitation (IP): Facilitates the enrichment and isolation of TagRFP-tagged proteins (cancer.gov).
3. Compatibility with Various Detection Methods
- Works with chemiluminescent, fluorescent, and enzyme-linked detection systems (ncbi.nlm.nih.gov).
- Can be used in conjunction with secondary antibodies for enhanced signal detection (nih.gov).
Applications in Research
1. Cellular Imaging and Localization Studies
The antibody enables high-resolution imaging of proteins tagged with TagRFP, helping researchers study protein dynamics, organelle targeting, and cellular trafficking (nih.gov).
2. Protein-Protein Interaction Studies
Using immunoprecipitation, scientists can isolate TagRFP fusion proteins and their interacting partners for downstream analysis (genome.gov).
3. Neuroscience and Developmental Biology
TagRFP is commonly used in neuroscience to track neuronal development and synaptic activity, with its antibody facilitating detailed studies in brain tissue (ninds.nih.gov).
4. Cancer Research
Researchers use TagRFP-tagged proteins in tumor progression studies, making this antibody valuable for cancer biomarker identification (cancer.gov).
Comparison with Other TagRFP Antibodies
| Feature | Goat Anti-TagRFP Polyclonal IgG | Rabbit Anti-TagRFP | Mouse Monoclonal Anti-TagRFP |
|---|---|---|---|
| Host Species | Goat | Rabbit | Mouse |
| Polyclonal/Monoclonal | Polyclonal | Polyclonal | Monoclonal |
| Applications | WB, IF, FACS, IP | WB, IF, IP | WB, IF |
| Sensitivity | High | Medium | High |
How to Use Goat Anti-TagRFP Polyclonal IgG Antibody
1. Western Blot Protocol
- Prepare cell lysates expressing TagRFP-fusion proteins.
- Separate proteins via SDS-PAGE (nih.gov).
- Transfer to a PVDF or nitrocellulose membrane.
- Incubate with Goat Anti-TagRFP Polyclonal IgG Antibody.
- Detect using a secondary anti-goat antibody conjugated to HRP or fluorescence.
2. Immunofluorescence Protocol
- Fix cells with paraformaldehyde.
- Permeabilize using Triton X-100.
- Block non-specific binding with BSA.
- Incubate with Goat Anti-TagRFP Antibody and a suitable secondary antibody.
- Image under a fluorescence microscope (nih.gov).
Troubleshooting Tips
1. Weak Signal in Western Blot?
- Optimize antibody concentration and incubation time (nih.gov).
2. High Background in Immunofluorescence?
- Increase blocking time and reduce antibody dilution (ncbi.nlm.nih.gov).
3. Poor Immunoprecipitation Yield?
- Ensure fresh lysates and use sufficient beads for antibody binding (cancer.gov).
Conclusion
The Goat Anti-TagRFP Polyclonal IgG Antibody is a powerful tool for detecting TagRFP-tagged proteins in a variety of research applications. With high specificity, strong signal detection, and broad usability, it serves as a key reagent in molecular and cellular biology studies. For further resources, visit NIH, NCBI, and Cancer.gov for the latest research and protocols (nih.gov).