GFP Tag Antibodies

Structure of GFP Tag

The GFP Tag is a relatively small protein with a molecular weight of approximately 27 kDa. This protein is composed of 238 amino acids and has a unique β-barrel-shaped three-dimensional structure, which encloses a chromophore. It can emit green fluorescence upon blue light excitation without the need for auxiliary factors. The chromophore is formed by the self-cyclization of the serine-threonine-glycine tripeptide at positions 65-67. This spontaneous fluorescence mechanism gives it a unique advantage in live cell imaging. The secondary structure of the GFP Tag is mainly composed of 11 β-sheet layers, which are arranged in a circular shape and form a highly stable cylindrical structure. The ends are closed by short-chain α-helices. This "β-barrel" conformation endows the protein with excellent chemical stability and thermal stability.

Fig. 1 The GFP tag has enzymatic activity and can cleave the CD81 protein into functional fragments.¹

Key structural features of the GFP Tag:

• Unique β-barrel structure
• The chromophore group is located at the center of the barrel
• The tripeptide sequence spontaneously forms the fluorescent chromophore
• Histidine is involved in chromophore maturation and proton transfer

Functions of GFP Tag

The main function of the GFP Tag is to serve as a fluorescent marker for tracking the expression location of fusion proteins within cells. However, it is also widely used in various biological researches, such as gene expression analysis, protein dynamic changes, and cell lineage tracing.

The fluorescence excitation spectrum of GFP Tag shows a main peak at 488 nm and an additional peak at 395 nm. The peak of the emission spectrum is at 509 nm. Its unique chromophore can spontaneously form and emit stable fluorescence without the need for additional substrates or cofactors, which gives it unique advantages in live-cell imaging and is widely applied in various fields ranging from molecular biology to developmental biology.

Applications of GFP Tag and GFP Tag Antibody in Literature

1. Carrillo Sanchez, Braulio, Matthew Hinchliffe, and Daniel G. Bracewell. "GFP‐tagging of extracellular vesicles for rapid process development." Biotechnology Journal 17.6 (2022): 2100583. https://doi.org/10.1002/biot.202100583

To accelerate the development of the extracellular vesicle (EV) process, this study designed stable CHO cells that stably express CD81-GFP. By leveraging the fluorescence characteristics of the GFP tag, rapid quantitative monitoring of EV concentration, purification loss, and chromatographic elution was achieved. This tool simplifies the analysis process of the EV purification procedure, facilitating the acceleration of process development.

2. Weill, Uri, et al. "Assessment of GFP tag position on protein localization and growth fitness in yeast." Journal of molecular biology 431.3 (2019): 636-641. https://doi.org/10.1016/j.jmb.2018.12.004

To investigate the impact of GFP tags on protein functions, this study systematically compared the differences between N-terminal and C-terminal fused GFP in yeast. Through competitive experiments, the adaptability of the two labeled strains was evaluated, and the physiological localization and the optimal labeling position of each protein were determined. This research provides a systematic reference for the precise use of protein tags.

3. Huet, Simon, et al. "Use of the nanofitin alternative scaffold as a GFP-ready fusion tag." PLoS One 10.11 (2015): e0142304. https://doi.org/10.1371/journal.pone.0142304

Research has developed a "ready-to-use GFP label", made up of GFP Nanofitin resistance. This tag can highly affinity bind to GFP and its spectral variants, and has excellent stability. The TNFα fused with this tag retains its full activity and can be flexibly labeled and detected by combining with different GFPs, providing a new tool for protein engineering.

4. Stevens, James C., et al. "Modification of superoxide dismutase 1 (SOD1) properties by a GFP tag–implications for research into amyotrophic lateral sclerosis (ALS)." PloS one 5.3 (2010): e9541. https://doi.org/10.1371/journal.pone.0009541

The study found that the GFP tag does not affect the distribution and movement of SOD1 within the cell, but it does alter its stability and enzymatic activity. Some fusion proteins undergo degradation. The results indicate that the GFP tag is suitable for localization studies, but caution is needed when analyzing the biochemical properties of the protein.

5. Rourke, Christine K., et al. "Endogenous localization of TOP-2 in C. elegans using a C-terminal GFP-tag." microPublication Biology 2021 (2021): 10-17912. https://doi.org/10.17912/micropub.biology.000402

For the study of dynamic positioning in the TOP - 2 in meiosis, the use of CRISPR/Cas9 constructed endogenous C side TOP - 2: : GFP nematode strains. The GFP-labeled TOP-2 exhibited a similar localization pattern to that of FLAG-labeled TOP-2, and did not significantly affect embryonic survival rate or sex chromosome segregation. The results indicate that this fusion protein is fully functional and suitable for in vivo localization studies.

Creative Biolabs: GFP Tag Antibodies for Research

Creative Biolabs specializes in the production of high-quality GFP Tag antibodies for research and industrial applications. Our portfolio includes monoclonal and polyclonal antibodies tailored for ELISA, Flow Cytometry, Western blot, immunohistochemistry, and other diagnostic methodologies.

• Custom GFP Tag Antibody Development: Tailor-made solutions to meet specific research requirements.
• Bulk Production: Large-scale antibody manufacturing for industry partners.
• Technical Support: Expert consultation for protocol optimization and troubleshooting.
• Aliquoting Services: Conveniently sized aliquots for long-term storage and consistent experimental outcomes.

For more details on our GFP Tag antibodies, custom preparations, or technical support, contact us at email.