In the realm of biochemical research and protein purification, the 3xFLAG peptide has become an indispensable tool for scientists. As someone who has spent significant time in the lab working with protein expression and affinity chromatography, 3X FLAG® Peptide - Sigma-Aldrich I have found that understanding the precise 3xflag peptide size and structural characteristics is essential for successful experimental outcomes.
Defining the Entity: What is the 3xFLAG Peptide?
The 3xFLAG peptide is a synthetic, hydrophilic 23-mer peptide designed to provide high-affinity binding for purification applications. Unlike the original single FLAG tag, the 3xFLAG version incorporates three repeated Asp-Tyr-Lys-Xaa-Xaa-Asp motifs. This increased length significantly enhances the binding affinity of the 3x FLAG tag to anti-FLAG antibodies, making it a superior choice for competitive elution.
When discussing the molecular weight of this construct, it is generally cited as approximately 2,861.87 Da (or roughly 2.9 kDa). Its small size is one of its most advantageous features. Because it is a compact peptide, it minimizes interference with the tertiary structure and biological function of the target protein, which is critical during sensitive procedures like protein crystallization.
Practical Considerations: Size and Usage
When sourcing this reagent, you will encounter various product formats. Whether you are looking for 10 mg quantities for large-scale purification or smaller aliquots like Size: 1.0 mg or Size, 4mg, 25mg, the quality of the peptides remains consistent. For those 3× FLAG peptide | Protein purification managing bulk inventory, you might encounter options like 3x FLAG peptide (25 mg; 5 x 5 mg), which allows for better stability by keeping unused portions stored at -20°C.
The 3xFLAG peptide (often referred to in literature as 3X FLAG peptide or simply 3xFlag) is typically provided as a lyophilized powder. When reconstituting, I have found that following the specific supplier’s protocol—often involving a buffer like TBS—ensures the highest recovery rate. The 3X FLAG peptides are highly specific for competitive elution, meaning they can effectively displace tagged proteins from affinity resins such as the ANTI-FLAG M2 gel.
Key Technical Specifications
* Sequence: The 3X FLAG peptide sequence is characterized by the MDYKD 3X FLAG Peptide HDGDYKDHDIDYKDDDDK motif.
* Molecular Weight: Approximately 2,862 g/mol.
* Purity: Typically >95% or >96% as determined by HPLC.
* Application: Ideal for SDS-PAGE, immunoprecipitation (IP), and protein identification.
Why the 3xFLAG Peptide Matters
In my own research, the shift from single to triple flags was a game-changer. The 3X FLAG peptide provides a much cleaner elution profile. Because of its small size, it does not easily aggregate, and its hydrophilic nature ensures that it remains soluble during the purification process. Whet 3xFLAG™Peptide; Empirical Formula (Hill Notation):. C120H169N31O49S ;Molecular Weight: 2861.87 ; UNSPSC Code: 12352202 ; NACRES: NA.32 ; Product Name.3X FLAG®Read more her you are working with 3X FLAG peptide (TFA) formulations or We do SPPS in our lab. 3xFlag is pretty large for SPPS at 22aa, and the aspartic acid residues present a challenge because of aspartimide Read more standard salt forms, the consistency of these peptides allows for highly reproducible resu The3x Flag-tag syntheticpeptidecontains 23 amino acid residue. Sequence: H-MDYKD HDGDY KDHDI DYKDD DDK-OH Eight amino acids at the C-terminus make up the Read more lts in western blotting and downstream protein analysis.
Final Thoughts on Selection
Choosing the right Size for your 3X Flag requirements depends on your throughput. For routine lab work, I usually keep a few vials of 3xFLAG peptide on hand. Always check the certificate of analysis for the molecular weight and ensure the storage conditions are strictly maintained at -20°C to preserve the integrity of the 3xFLAG construct. By respecting these technical par 3X FLAG peptide | Protein Marker ameters, you can ensure that your protein purification workflows remain efficient and reliable.