Antigenic peptides of M3R

Antigenic peptides of M3R

• COA/MS/HPLC Report
• Handling and Storage of Synthetic Peptides
• Material Safety Data Sheets (MSDS)

Batch to batch variation of the purity

1 Vial

H-VLVNTFCDSCIPKTYWNLGY-OH

H-Val-Leu-Val-Asn-Thr-Phe-Cys-Asp-Ser-Cys-Ile-Pro-Lys-Thr-Tyr-Trp-Asn-Leu-Gly-Tyr-OH

20

95.38%

C₁₀₈H₁₅₈N₂₄O₃₀S₂

2336.67

Synthetic

Serum levels of IgG against Sjögren’s syndrome-related antigen A (SSA) and M3 muscarinic receptor (M3R) were examined by a standard sandwich enzyme-linked immunosorbent assay (ELISA). Briefly, 96-well MaxiSorp plates were coated with antigen peptides (5 μg/ml) at 4 °C overnight. Plates were washed and incubated with blocking buffer (0.5% gelatin, 0.5% bovine serum albumin and 0.05% Tween 20 in PBS) at room temperature for 1 h. Serum samples were diluted (1:100) and incubated in plates for 2 h at room temperature, followed by incubation of biotin-conjugated anti-mouse IgG (BioLegend, 0.5 μg/ml) for 1 h. After washing, HRP Streptavidin (BioLegend, 1:1000) was added, and the plates were incubated for 30 min. Then, plates were washed and freshly prepared TMB substrate (BioLegend, 50 μl) was added. After 10 min, 20 μl stop solution (3 M H2SO4) was added and absorbance at 450 nm was measured using a Sunrise microplate reader (Tecan, Männedorf, Switzerland). Antigenic peptides of SSA (AVALREYRKKMDIPA) and M3R (VLVNTFCDSCIPKTYWNLGY).

Normally, this peptide will be delivered in lyophilized form and should be stored in a freezer at or below -20 °C. For more details, please refer to the manual:Handling and Storage of Synthetic Peptides

• Xiao F, Lin X, Tian J, Wang X, Chen Q, Rui K, Ma J, Wang S, Wang Q, Wang X, Liu D, Sun L, Lu L. Proteasome inhibition suppresses Th17 cell generation and ameliorates autoimmune development in experimental Sjögren's syndrome. Cell Mol Immunol. 2017 Jul 10;14(11):924–34. doi: 10.1038/cmi.2017.8. Epub ahead of print. PMID: 28690324; PMCID: PMC5675963.

Trifluoroacetic acid (TFA) is a strong acid, which is commonly used to cleave synthesized peptides from solid-phase resins and is also used to improve HPLC performance in the peptide purification step. By default, custom peptides are delivered as lyophilized TFA salts, and can contain as much as 10-45% TFA.

TFA in custom peptides can cause inexplicable discrepancies in subsequent assay data. For instance, TFA in nM concentrations has been shown to interfere with cellular assays, inhibiting cellular proliferation in some instances, and increasing cell viability in others . It has also been found to be an unintended allosteric modulator of the glycine receptor, GlyR.

TFA Removal Service is recommended for:

• Peptides that will be used in cellular assays
• Peptides that will be used as APIs or in manufactured products
• For hydrophilic peptides containing numerous basic residues