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Siinfekl peptide

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SIINFEKL peptide is a synthetic peptide that consists of the amino acid sequence SIINFEKL. It is commonly used in experimental research applications, particularly in the field of immunology and cell biology.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Brefeldin a and monensin, by BioLegend (1 mentions) Brefeldin a, by BioLegend (1 mentions) Monensin, by BioLegend (1 mentions) Naive cd8 t cell isolation kit, by Miltenyi Biotec (1 mentions) 6 well plate, by Corning (1 mentions) Triton x 100, by PerkinElmer (1 mentions) Glutaraldehyde, by Electron Microscopy Sciences (1 mentions) Easysep mouse na ve cd8 t cell isolation kit, by STEMCELL (1 mentions) Mojosort mouse anti apc nanobeads, by BioLegend (1 mentions) Atp detection assay kit luminescence, by Cayman Chemical (1 mentions) Recombinant murine il 15, by Thermo Fisher Scientific (1 mentions) Poly d lysine, by Merck Group (1 mentions) Seahorse xf cell mito stress test kit, by Agilent Technologies (1 mentions)

5 protocols using siinfekl peptide

1

Vac-OVA Infection and LPS Treatment in Infant Mice

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Fifteen-day-old CTRL and MAT C57BL/6J infant mice were infected with Vac-OVA (1 × 104 PFU i.p.) and orally treated with E. coli 0111:B4-derived LPS (50 μg orogastric; InvivoGen) beginning on the day of the infection and continuing every other day for 10 days. Mice were monitored daily for weight loss and appearance of illness. Eleven days after infection, mice were euthanized by CO2 inhalation. Spleens were mechanically disrupted to obtain single cell suspensions and then treated with ACK buffer to lyse red blood cells. For the detection of cytokines, the splenocytes were cultured for 5 h in RPMI-10 with SIINFEKL peptide (5 μM; New England Peptide), phorbol 12-myristate 13-acetate (PMA) (10 ng/ml), and ionomycin (1 μg/ml) in the presence of brefeldin A and monensin (BioLegend).
Gonzalez-Perez G, & Lamousé-Smith E.S. (2017). Gastrointestinal Microbiome Dysbiosis in Infant Mice Alters Peripheral CD8+ T Cell Receptor Signaling. Frontiers in Immunology, 8, 265.
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2

Adoptive Transfer and Vaccinia Infection

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Control and MAT OT-I CD8+ T cells pooled from littermates (1.5 × 105/100 μl PBS) were transferred into age- and gender-matched CTRL Rag1KO recipients by intraperitoneal (i.p.) injection. Twenty-four hours after OT-I adoptive cell transfer, recipient mice were infected i.p. with 1 × 104 PFU of recombinant vaccinia-ovalbumin (Vac-OVA) by i.p. injection. Mice were monitored daily for weight loss and appearance of illness. Eight days after infection, mice were euthanized by CO2 inhalation. Peritoneal exudate cells (PEC) were aspirated following lavage of the peritoneum with 1 ml sterile PBS. Spleens and mesenteric lymph nodes (MLN) were mechanically disrupted to obtain single cell suspensions and then treated with ACK buffer to lyse red blood cells. For the detection of cytokines, the cells were cultured for 5 h in RPMI-10 with SIINFEKL peptide (5 μM; New England Peptide) in the presence of brefeldin A and monensin (BioLegend).
Gonzalez-Perez G, & Lamousé-Smith E.S. (2017). Gastrointestinal Microbiome Dysbiosis in Infant Mice Alters Peripheral CD8+ T Cell Receptor Signaling. Frontiers in Immunology, 8, 265.
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3

Adoptive Transfer of Activated OT-I Cells

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Naive CD8+ T cells were purified from LN and spleen single-cell suspensions by immunomagnetic negative cell selection using the Miltenyi naive CD8+ T cell isolation kit and adoptively transferred into sex-matched recipients by retro-orbital injection. For the in vitro activation of T cells, OT-I × Tcra−/− splenocytes were pulsed with 1 μM SIINFEKL peptide (New England Peptide) in 1 ml of T cell medium (RPMI, 10% FCS, 1% HEPES, 1% sodium pyruvate, 1% GlutaMAX, 1% non-essential amino acids, 55 μM 2-mercaptoethanol) for 1 h at 37 °C, diluted in 9 ml of T cell medium, and cultured at 37 °C in 5% CO2. Two days later, 20 ng/ml of IL-2 was added and cell density was maintained at 106 cells/ml. OT-I cells were adoptively transferred on day 5 after activation by retro-orbital injection.
For T cell depletion, one dose of anti-Thy1.2 mAbs (3 μg, clone 30H12) was injected i.v.
Mani V., Bromley S.K., Äijö T., Mora-Buch R., Carrizosa E., Warner R.D., Hamze M., Sen D.R., Chasse A.Y., Lorant A., Griffith J.W., Rahimi R.A., McEntee C.P., Jeffrey K.L., Marangoni F., Travis M.A., Lacy-Hulbert A., Luster A.D, & Mempel T.R. (2019). Migratory DCs activate TGF-β to precondition naive CD8+ T cells for tissue-resident memory fate. Science (New York, N.Y.), 366(6462), eaav5728.
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4

OT-1 T cell Priming Protocol

Cited 2 times
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DsRed fluorescent protein-expressing OT–1 mice (C57BL/6–TgTcra/Tcrb homozygous, 1100 Mjb/J) were a generous gift from Professor Mei Wu (Wellman Center for Photomedicine, Massachusetts General Hospital and Harvard Medical School, Boston). Mice 10 weeks of age or older were euthanized by CO2 asphyxiation following an approved IACUC protocol. Spleens were collected aseptically immediately after euthanasia and T cells were primed following a published protocol (39 (link)) with minor modifications. Briefly, red blood cells in the splenic suspension were depleted by incubating with ammonium–chloride–potassium lysing buffer (Gibco, A1049201) for 4 minutes. Three days before addition to 3D cultures, T cell blasts were generated in 6–well plates (Corning, 353046) by inoculating 0.75 μg/mL of SIINFEKL peptide (ovalbumin sequence 257–264, New England Peptide, BP10–915) into 1×106 splenocytes/mL for 3 days in the presence of 10 U/ml recombinant murine interleukin–2 (IL–2) cytokine (Peprotech Inc., 212–12). This protocol yields a >90% pure solution of CD8+ T cells (39 (link), 40 (link)) and was not purified further. Cells were washed twice in RPMI 1640 media before addition to 3D cultures. In all cases, T cells were maintained in SuppRPMI medium further supplemented with 50 μM β–mercaptoethanol (Sigma, M3148) and 10 U/mL recombinant murine IL–2 (T cell RPMI).
Kercher E.M., Nath S., Rizvi I, & Spring B.Q. (2019). Cancer Cell-targeted and Activatable Photoimmunotherapy Spares T Cells in a 3D Co-culture Model. Photochemistry and photobiology, 96(2), 295-300.
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5

Murine CD8+ T Cell Isolation and Metabolic Profiling

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EasySep Mouse naïve CD8 T cell isolation kits (Stemcell Technologies cat:19858A); Mojosort mouse anti-APC nanobeads (Biolegend Cat:480072); ATP detection assay kit-luminescence (Cayman Chemical cat:700410); DAPI (Thermo Fisher cat:D1306); Seahorse XF Cell Mito Stress Test Kit (Seahorse Agilent cat:103015-100); 2-DG (Cayman Chemical cat:14325); SIINFEKL peptide (New England peptide Lot:V1355-37/40); recombinant human IL-2 (TECIN cat:Ro23-6019); recombinant murine IL-15 (PeproTech cat:210-15); poly-D-lysine (Millipore Sigma cat:P6407); Glutaraldehyde (Electron Microscopy Science cat:16000); NaBH4 (Alfa Aesar stock#:35788); Triton X-100 (PerkinElmer cat:N9300260).
Sun Y., Preiss N.K., Valenteros K.B., Kamal Y., Usherwood Y.K., Frost H.R, & Usherwood E.J. (2020). Zbtb20 restrains CD8 T cell immunometabolism and restricts memory differentiation and anti-tumor immunity. Journal of immunology (Baltimore, Md. : 1950), 205(10), 2649-2666.
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