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15 mer peptides

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15-mer peptides are synthetic peptides consisting of 15 amino acids. They are a versatile tool for various applications in biochemistry and molecular biology research.

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Lab products found in correlation

Brefeldin a, by Merck Group (2 mentions) Anti cd28, by BD (2 mentions) Ifn γ b27, by BD (2 mentions) Tnf mab11, by Thermo Fisher Scientific (2 mentions) Cd69 fn50, by BD (2 mentions) Anti cd49d co stimulatory mab, by BD (2 mentions) Penicillin streptomycin antibiotic, by Merck Group (1 mentions) Hepes, by Merck Group (1 mentions)

4 protocols using 15 mer peptides

1

Quantification of SIV-specific CD8+ T Cells

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SIV-specific CD8+ T cells were quantified in mononuclear cells isolated from blood and LN by flow cytometric intracellular cytokine analysis, as previously described in detail45 (link). Briefly, mixes of sequential (11 amino acid overlapping) 15-mer peptides (AnaSpec) spanning the SIVmac239 Gag, Env, Pol, Nef, Rev/Tat, Vif/Vpr/Vpx proteins were used as antigens in conjunction with anti-CD28 (BD Biosciences) and anti-CD49d co-stimulatory mAb (BD Biosciences). Cells were incubated at 37 °C with peptide mixes/pools and co-stimulatory antibodies for 1 hr, followed by an additional 8 hr. incubation in the presence of Brefeldin A (5 μg/ml, Sigma-Aldrich). Co-stimulation in the absence of peptides served as background control. Cells were stained with fluorochrome-conjugated monoclonal antibodies [CD3 (SP34-2), CD4 (L200), CD8a (SK1), IFN-γ (B27)(BD Biosciences), TNF (MAB11, eBioscience), CD69 (FN50, BD Biosciences)], and data were collected on an LSRII (BD Biosciences) and analyzed using the FlowJo software program (version 8.8.7; Tree Star, Inc.). Response frequencies to each peptide mix were defined by the intracellular expression of TNF and/or IFN-γ by CD3+, CD4−, CD8+ T cells after subtraction of background. Response frequencies to each peptide mix were added to obtain total SIV-specific response frequencies.
Fukazawa Y., Lum R., Okoye A.A., Park H., Matsuda K., Bae J.Y., Hagen S.I., Shoemaker R., Deleage C., Lucero C., Morcock D., Swanson T., Legasse A.W., Axthelm M.K., Hesselgesser J., Geleziunas R., Hirsch V.M., Edlefsen P.T., Piatak M J.r., Estes J.D., Lifson J.D, & Picker L.J. (2015). A B cell follicle sanctuary permits persistent productive SIV infection in elite controllers. Nature medicine, 21(2), 132-139.
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2

Quantification of SIV-specific CD8+ T Cells

Cited 1 time
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SIV-specific CD8+ T cells were quantified in mononuclear cells isolated from blood and LN by flow cytometric intracellular cytokine analysis, as previously described in detail45 (link). Briefly, mixes of sequential (11 amino acid overlapping) 15-mer peptides (AnaSpec) spanning the SIVmac239 Gag, Env, Pol, Nef, Rev/Tat, Vif/Vpr/Vpx proteins were used as antigens in conjunction with anti-CD28 (BD Biosciences) and anti-CD49d co-stimulatory mAb (BD Biosciences). Cells were incubated at 37 °C with peptide mixes/pools and co-stimulatory antibodies for 1 hr, followed by an additional 8 hr. incubation in the presence of Brefeldin A (5 μg/ml, Sigma-Aldrich). Co-stimulation in the absence of peptides served as background control. Cells were stained with fluorochrome-conjugated monoclonal antibodies [CD3 (SP34-2), CD4 (L200), CD8a (SK1), IFN-γ (B27)(BD Biosciences), TNF (MAB11, eBioscience), CD69 (FN50, BD Biosciences)], and data were collected on an LSRII (BD Biosciences) and analyzed using the FlowJo software program (version 8.8.7; Tree Star, Inc.). Response frequencies to each peptide mix were defined by the intracellular expression of TNF and/or IFN-γ by CD3+, CD4−, CD8+ T cells after subtraction of background. Response frequencies to each peptide mix were added to obtain total SIV-specific response frequencies.
Fukazawa Y., Lum R., Okoye A.A., Park H., Matsuda K., Bae J.Y., Hagen S.I., Shoemaker R., Deleage C., Lucero C., Morcock D., Swanson T., Legasse A.W., Axthelm M.K., Hesselgesser J., Geleziunas R., Hirsch V.M., Edlefsen P.T., Piatak M J.r., Estes J.D., Lifson J.D, & Picker L.J. (2015). A B cell follicle sanctuary permits persistent productive SIV infection in elite controllers. Nature medicine, 21(2), 132-139.
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3

Peptide Pool Preparation for HIV Immunogenicity Assays

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HIVconsv-derived 15-mer peptides overlapping by 11 aa (Ana Spec, San Jose, CA 95131) were reconstituted to 40 mg/ml in dimethylsulfoxide (DMSO) and diluted to working stock solutions of 4 mg/ml in phosphate-buffered saline. For ELISPOT assays, peptides were combined into six pools, P1–P6, of 32–36 peptides per pool in R10 (RPMI 1460 supplemented with 10% FBS, 2 mmol/l L-glutamine, 1 mmol/l sodium pyruvate, 10 mmol/l N-2-hydroxyethylpiperazine-N9-2-ethanesufonic acid (HEPES) and penicillin-streptomycin antibiotics; Sigma Aldrich, St. Louis, MO) and 2×-concentrated stocks of 3 µg/ml were prepared and aliquoted into peptide plates along with the positive and negative controls. For frozen ELISPOT assay, peptides were arranged into seven pools comprising nonjunctional peptides in P1–P6 and junctional peptides in JXN. For identification of stimulatory 15-mers, peptides were arranged into 3-D matrix pools. For ICS studies, the peptides were combined into four pools covering Gag, Pol (two pools), and Env+Vif. A pool of FEC (“Flu”, EBV and CMV) peptides consisting of 32 previously defined CD8+ T-cell epitopes from influenza virus, Epstein-Barr virus, and cytomegalovirus (CEF; NIH AIDS Research and Reference Program) was reconstituted in dimethylsulfoxide and used at a final concentration of 1 µg/ml as a positive control. All peptide stocks and plates were stored at −80oC until use.
Mutua G., Farah B., Langat R., Indangasi J., Ogola S., Onsembe B., Kopycinski J.T., Hayes P., Borthwick N.J., Ashraf A., Dally L., Barin B., Tillander A., Gilmour J., De Bont J., Crook A., Hannaman D., Cox J.H., Anzala O., Fast P.E., Reilly M., Chinyenze K., Jaoko W., Hanke T, & HIV-CORE 004 study group T. (2016). Broad HIV-1 inhibition in vitro by vaccine-elicited CD8+ T cells in African adults. Molecular Therapy. Methods & Clinical Development, 3, 16061-.
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4

Reconstitution of HIVconsv Peptides

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HIVconsv-derived 15-mer peptides overlapping by 11 amino acids and spanning the entire HIVconsv protein (Ana-Spec, San Jose, USA) were reconstituted to 10–40 mg/ml in DMSO and diluted to working stock solutions of 4 mg/ml in PBS as described previously [15 (link)].
Borthwick N.J., Lane T., Moyo N., Crook A., Shim J.M., Baines I., Wee E.G., Hawkins P.N., Gillmore J.D., Hanke T, & Pepys M.B. (2018). Randomized phase I trial HIV-CORE 003: Depletion of serum amyloid P component and immunogenicity of DNA vaccination against HIV-1. PLoS ONE, 13(5), e0197299.
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