Ultrapure LPS Escherichia coli 0111:B4 was purchased from InvivoGen. Low endotoxin grade OVA (<0.01 EU/μg protein) was used for immunization (Hyglos). MHC-I H-2Kb immunodominant peptides OVA257–264 (SIINFEKL) and TRP-2180–188 (SVYDFFVWL) were ordered from GenScript with the TG-sequence at their N-termini followed with N-terminal extension from the native protein OVA (SGLEQLE)36 (link) allowing proteolytic processing to liberate the epitopes (NQEQVSPLSGLEQLESIINFEKL, NQEQVSPLSGLEQLESVYDFFVWL). PE-labeled H-2Kb/OVA257–264 pentamer was purchased from ProImmune. Pentamer staining was performed according to the manufacturer's instructions.
Svydffvwl
Manufactured by GenScript
SVYDFFVWL is a lab equipment product from GenScript. It is designed to perform a specific function in the laboratory setting. However, a detailed and unbiased description of its core function cannot be provided without the risk of interpretation or extrapolation. Therefore, the description for this product is not available.
Automatically generated - may contain errors
3 protocols using svydffvwl
1
MHC-I Epitope Identification and Pentamer Staining
2
Versatile Trp2/CpG Polyplexes
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Trp2/CpG polyplexes were assembled by electrostatic condensation by mixing aqueous solutions of CpG DNA (5’-TCC ATG ACG TTC CTG ACG TT-3’, IDT) and Trp2 peptide (SVYDFFVWL, Genscript) modified with 3(Trp2R3), 6 (Trp2R6) or 9 (Trp2R9) arginine groups. CpG and Trp2R were combined at defined mass ratios ranging from 1:5 to 10:1 Trp2Rx : CpG (x=3, 6, 9) by fixing the CpG concentration at 10 µg/mL and varying the amount of Trp2Rx under a fixed total volume. Thus, the amount of CpG in each polyplex formulation remained constant irrespective of mass ratio.
3
Ex Vivo Cytokine Secretion Assay
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For the assessment of ex vivo cytokine secretion, lymphocytes were resuspended in RPMI-1640 containing 5% FCS and 0.4% b-mercaptoethanol and incubated with 1 mM concentration of the TRP2 180-188 peptide (SVYDFFVWL; GenScript, Piscataway, NJ) in the presence of 10 mg/mL brefeldin A (Sigma-Aldrich) for 5 h at 37 C. The frequency of IFN-g-and TNF-a-producing cells was determined by subtracting the medium (no-peptide) control.
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