Anti-RFP/mCherry Agarose Beads: The Optimal Choice for Purifying Fluorescently Tagged Proteins

Introduction

Protein tags are peptides or proteins fused to target proteins to facilitate their expression, detection, tracing, and purification. Common protein tags are categorized based on their applications, including peptide tags (e.g., Flag tag, His tag), affinity tags (e.g., MBP, GST), and fluorescent tags (e.g., mCherry).

Tag antibodies are specific antibodies that recognize and bind to particular tag sequences on fusion proteins, enabling the detection and purification of target proteins. They are widely used in protein expression, localization, interaction studies, and functional research. When conjugated to agarose beads, tag antibodies can efficiently pull down target proteins from cell lysates, facilitating protein isolation and purification, commonly applied in immunoprecipitation and protein purification.

mCherry

mCherry is one of the most widely used red fluorescent proteins (RFPs) in biological research. It is a monomeric red fluorescent protein derived from the Discosoma sp. red fluorescent protein DsRed through genetic engineering. mCherry has maximum absorption and emission wavelengths at approximately 587 nm and 610 nm, respectively. When fused to target proteins, mCherry's fluorescence activity and the target protein's function remain independent without significant mutual interference.

Advantages of mCherry

• High Fluorescence Brightness: Exhibits excellent fluorescence properties in the red spectrum, providing strong signals during imaging.

• Superior Photostability: Demonstrates strong resistance to photobleaching, maintaining stable fluorescence signals even under prolonged illumination.

• Low Background Fluorescence: Offers clear and distinct fluorescence labeling in experiments, facilitating cell observation and counting.

• Rapid Maturation Time: With a t0.5 of only 15 minutes, it is suitable for rapid responses in promoter activity reporter systems.

• Monomeric Form: Does not require other subunits to form an active structure, ensuring that fusion does not disrupt its functional integrity.

• Suitable for Live-Cell Imaging: mCherry is ideal for live-cell imaging, enabling the study of dynamic changes in gene expression.

• Broad Adaptability: Widely used in both prokaryotic and eukaryotic cells, suitable for various biological experiments, including promoter activity studies, fluorescence resonance energy transfer (FRET), and other quantitative assays.

• Ease of Operation: The mCherry expression system does not require inducers, making it suitable for scenarios where inducer addition is challenging, such as recombinant bacteria expressing proteins in the gastrointestinal environment of experimental animals.

• Compatibility with Other Fluorescent Proteins or Reporter Genes: Enables multicolor labeling and co-localization studies, and can be combined with CRISPR activation systems to monitor endogenous gene expression.

The numerous advantages of mCherry, including high brightness, rapid maturation, excellent spectral properties, monomeric form, low background interference, suitability for live-cell imaging, compatibility with other systems, broad applicability, and simplified operational procedures, make it a powerful tool for studying gene expression and cellular biological functions.

S-RMab® RFP/mCherry Agarose Beads

Starter introduces Rabbit Anti-RFP/mCherry Agarose Beads, which utilize a 4% agarose gel matrix to achieve high-affinity binding with mCherry protein.

• Efficient Capture: Each 25 µL of agarose can bind up to 15 µg of RFP fusion protein.

• Versatile Applications: Purification of RFP-tagged fusion proteins and immunoprecipitation.

• Ready-to-Use: Available as off-the-shelf antibodies for immediate use.

Product Data Demonstration

Rabbit Anti-RFP/mCherry Agarose Beads

• Lane 1: Input

• Lane 2: Flow-through

• Lane 3: RFP fusion protein

mCherry Recombinant Rabbit mAb

Western Blot Results

• Lane 1: 293T cell lysate

• Lane 2: 293T empty vector cell lysate

• Lane 3: 293T cells transfected with Histone H3-mCherry-E-tag