Cef peptides

Manufactured by Mabtech

Sourced in United States

CEF peptides are synthetic peptides designed for cell-based experiments. These peptides are used as positive and negative controls in various immunological assays, such as ELISpot and flow cytometry, to evaluate cellular immune responses.

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Lab products found in correlation

Cd4 pe, by BD (1 mentions) Cd45ra apc, by BD (1 mentions) Cd8 apc cy7, by BD (1 mentions) Siinfekl peptide, by Merck Group (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

CEF peptide pool (8 citations) CEF control peptide pool (3 citations)

4 protocols using cef peptides

Peptide-Specific Humoral and Cellular Immune Responses

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Humoral immune responses specific to each of the 31 peptide candidates were determined by peptide-specific IgG levels using the Luminex system (Luminex, Austin, TX, USA), as previously reported
[18 (link)]. If the titers of peptide-specific IgG to at least one of the vaccinated peptides in the post-vaccination plasma were more than two-fold higher than those in the pre-vaccination plasma, the changes were considered to be significant, as previously reported
[14 (link)-17 (link)]. Cellular immune responses specific to the vaccinated peptides were evaluated by interferon (INF)-γ ELISPOT using peripheral blood mononuclear cells (PBMCs) as previously reported
[14 (link)-17 (link)]. As a control, cellular immune responses specific to CEF peptides (MABTECH, Cincinnati, OH, USA), a mixture of virus-derived cytotoxic T lymphocyte (CTL) epitopes, were also examined.

Takahashi R., Toh U., Iwakuma N., Takenaka M., Otsuka H., Furukawa M., Fujii T., Seki N., Kawahara A., Kage M., Matsueda S., Akagi Y., Yamada A., Itoh K, & Sasada T. (2014). Feasibility study of personalized peptide vaccination for metastatic recurrent triple-negative breast cancer patients. Breast Cancer Research : BCR, 16(4), R70.

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Sorting and Characterizing Naive and Memory T Cells

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Cryopreserved PBMC samples were labelled using CD4-PE, CD8-APC-Cy7, CD45RA-APC (all Becton Dickinson) and CD62L-ECD (Beckman Coulter) on ice, following overnight rest to restore CD62L expression (Figure A in S1 File). CD4+ and CD8+ gated lymphocytes were sorted (Influx; Becton Dickinson) on the basis of CD45RA and CD62L expression [20 (link)]. Sort purities were regularly >98%. We confirmed naïve and memory populations by stimulating naïve or memory CD8+ T cells with cytomegalovirus (CMV), Epstein-Barr virus (EBV) and influenza derived peptides (CEF peptides; Mabtech) and performing an IFN-γ ELISpot assay as described above. CEF peptides responses were only detected in stimulated memory phenotype T cells, but not naïve populations (Figure B in S1 File).

Lucas A., Lucas M., Strhyn A., Keane N.M., McKinnon E., Pavlos R., Moran E.M., Meyer-Pannwitt V., Gaudieri S., D’Orsogna L., Kalams S., Ostrov D.A., Buus S., Peters B., Mallal S, & Phillips E. (2015). Abacavir-Reactive Memory T Cells Are Present in Drug Naïve Individuals. PLoS ONE, 10(2), e0117160.

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Measuring Humoral and CTL Immune Responses to Peptide Vaccination

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Humoral immune responses specific to each of the 32 peptide candidates were determined by measuring the levels of peptide-specific IgG and IgG subclasses (IgG1, IgG2, IgG3, and IgG4) using the Luminex system (Luminex, Austin, TX), as previously reported [10 (link)–15 (link)]. If the titers of peptide-specific IgG to at least one of the vaccinated peptides after the 8th vaccination were more than twofold higher than those before vaccination, the changes were considered to be significant, as previously reported [10 (link)–15 (link)]. CTL responses specific to the vaccinated peptides were evaluated by interferon- (IFN-) γ ELISPOT assay using peripheral blood mononuclear cells (PBMCs) before and after vaccination as previously reported [10 (link)–15 (link)]. As a control, CTL responses specific to CEF peptides (MABTECH, Cincinnati, OH), a mixture of virus-derived CTL epitopes, were also examined.

Yutani S., Ueshima K., Abe K., Ishiguro A., Eguchi J., Matsueda S., Komatsu N., Shichijo S., Yamada A., Itoh K., Sasada T., Kudo M, & Noguchi M. (2015). Phase II Study of Personalized Peptide Vaccination with Both a Hepatitis C Virus-Derived Peptide and Peptides from Tumor-Associated Antigens for the Treatment of HCV-Positive Advanced Hepatocellular Carcinoma Patients. Journal of Immunology Research, 2015, 473909.

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T-cell Assays with Inhibitors and PGE2

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Human T-cell assays were performed with peripheral blood mononuclear cell (PBMC) isolated from a fresh leukopak (catalog no. 70500.2, StemCell Technologies). Human PBMC were thawed in warm RPMI+ Benzonase, enriched for T cells using magnetic separation (catalog no. 130-096-535, Miltenyi Biotec), then plated at 2 × 10⁶ cells/mL in 100 μL RPMI supplemented with 10% FBS (catalog no. 10438026, Gibco) and penicillin-streptomycin (catalog no. 15140163, Gibco). Mouse T-cell activation assays were performed with unenriched single-cell suspensions isolated after red blood cell (RBC) lysis (catalog no. A1049201, Gibco) of pooled lymph nodes and spleens from OT-1 Mice [Jax, C57BL/6-Tg(TcraTcrb)1100 Mjb/J] and plated at 0.5–1 × 10⁶ cells per well. In both human and mouse assays, cells were then preincubated with EP inhibitors for 20 minutes at 37°C, followed by PGE2 (catalog no. P0409, Sigma-Aldrich) for 20 minutes at 37°C, exposed to varying concentrations of PGE2 (10–1,000 nmol/L) for 30 minutes at 37°C. After these preincubation steps, CEF peptides (catalog no. 3616-1, Mabtech) or SIINFEKL peptide (catalog no. S7951, Sigma-Aldrich) were added, and all components were incubated for 6 additional hours at 37°C. At that time, supernatants were collected, and cytokines were measured by Luminex (catalog no. HCYTA-60K-PX48, Millipore).

Francica B.J., Holtz A., Lopez J., Freund D., Chen A., Wang D., Powell D., Kipper F., Panigrahy D., Dubois R.N., Whiting C.C., Prasit P, & Dubensky T.W. (2023). Dual Blockade of EP2 and EP4 Signaling is Required for Optimal Immune Activation and Antitumor Activity Against Prostaglandin-Expressing Tumors. Cancer Research Communications, 3(8), 1486-1500.

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