In many protein research fields, biotin-labeled peptides are usually combined with avidin to be detected or purified: Western blot (WB), ELISA, immunoprecipitation (IP),
Peptide affinity purification, immunohistochemistry (IHC), cell surface labeling, flow cytometry/fluorescence activated cell classification (FACS).
Beike Nano provides biotin-labeled peptide synthesis services. Generally, biotin can be labeled at the N-terminus or C-terminus (via Lys). We recommend N-terminal tagging, and its tagging
The power is higher, the time required is shorter, and the operation is simpler.
Biotin can be separated from peptides by various linkers or spacers. It is generally recommended to introduce a spacer such as Ahx (a 6-carbon linker) to make biotin more stable
Or more flexible.
Beike Nano provides N-terminal or C-terminal biotin labels: Biotin (N terminus), Lys (Biotin) (middle), Lys (Biotin) (C terminus).
Biotinylation with linker (Ahx): Biotin-Ahx (N terminus), Lys(Ahx-Biotin)(middle), Lys(Ahx-Biotin)(C terminus).
Analysis of the Pull-down Experiment Scheme of Biotin-labeled Peptides
Biotin-labeled peptides can be conjugated with avidin magnetic beads to generate resin for peptide pulldown tests. First load the biotin-labeled peptide to Immobilized
Streptavidin beads (Pierce) are anchored on magnetic beads with avidin, and then incubated with cell lysate. Cells are lysed in 1% (v/v) Nonidet P-40,
150 mM sodium chloride, 50 mM Tris-HCl, pH 7.5, protease inhibitors (Complete Tablets, Roche Applied
Science), adding 1 mM sodium orthovanadate as a phosphatase inhibitor. The same amount of protein was incubated with the anchored peptide at 4℃ 6
hour. After extensive washing and boiling in the SDS sample buffer, the binding protein can be eluted from the anchored peptide, or, with 50 mM
dithiothreitol breaks the peptide and protein complex from the magnetic beads. Combine the eluates from the active peptide and control target peptide pull-downs for further
Step analysis.
The figure above is a schematic diagram of streptavidin-biotin bond strength measurement. Due to their unusually high affinity binding, the most prominent ligand-receptor pair is Streptomyces
Avidin-biotin (streptavidin-biotin) and avidin-biotin (avidin-biotin). Both of these proteins have a tetrameric structure,
In this way they can bind 4 ligands.
Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance |
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Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance.The website pictures are from the Internet. The pictures are for reference only. Please take the real object as the standard. In case of infringement, please contact us to delete them immediately. |