circRNA Pull-Down Kit (Animal)
$650.00 - $1,015.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: CPDKA-12 (for 12 reactions)
Cat. No.: CPDKA-24 (for 24 reactions)
Description
circRNA Pull-Down Kit utilizes the strong affinity between the "F2 tag" and its specific ligand to efficiently retrieve target circRNA and its associated proteins from cells. The F2 tag is a short RNA sequence that, when linked to the target circRNA, has minimal impact on its structure and function.
Before the experiment, the F2 tag sequence is ligated to the target circRNA sequence to construct an F2-circRNA overexpression vector. This vector is then transfected into cells, which are subsequently lysed. Specific ligand-coated magnetic beads are used to pull down the F2-circRNA and its bound proteins. After removing unbound proteins, various methods can be used to elute the proteins. The isolated protein products can be analyzed using Western blot to detect known proteins or mass spectrometry (LC-MS/MS) to identify unknown proteins.
Applications
Protein-protein interactions are the primary mode through which proteins exert their biological functions. Proteins perform various functions by binding to different partners, forming complex signaling networks. The pull-down kit can be applied to several research directions or fields, including:
- Determining whether two proteins have a direct interaction;
- Identifying the domains involved in protein-protein binding;
- Studying the mechanisms of pathogen invasion, such as viruses and bacteria, into host cells;
- Exploring the mechanisms of enzyme-substrate interactions;
- Investigating protein regulatory complexes.
Advantages
High Retrieval Efficiency: The strong affinity between the F2 tag sequence and the F2 ligand-coated magnetic beads enables efficient retrieval of F2-circRNA and its associated proteins.
No Sequence Limitation: The method is not restricted by the sequence at the circRNA junction, making it suitable for various circRNA sequences.
Minimal Impact: The F2 tag is very short, only 16 nt, which has minimal effect on the structure or function of the target RNA.
Compatible with Subsequent Experiments: The elution buffer is compatible with downstream Western Blot and mass spectrometry (LC-MS/MS) analyses.
Ease of Use: The procedure is straightforward, time-efficient, and suitable for beginners.
Brief Usage Procedure
Construct the F2-circRNA expression vector, transfect target cells, and perform expression detection.
Lysate the samples and prepare the F2 ligand-coated magnetic beads.
Add the cell lysate to the F2 ligand-coated magnetic beads and incubate at 4°C for 2-4 hours.
Wash and elute to obtain the target protein and its interacting protein complexes.
Only for research and not intended for treatment of humans or animals
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