Mouse il 2 duoset

Manufactured by R&D Systems

The Mouse IL-2 DuoSet is a laboratory tool used for the quantitative determination of mouse Interleukin-2 (IL-2) levels in cell culture supernatants, cell lysates, serum, and plasma. It provides the necessary components to develop an enzyme-linked immunosorbent assay (ELISA).

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Lab products found in correlation

Ancillary reagent kit 2, by R&D Systems (1 mentions) Balb c mice, by InvivoGen (1 mentions) Alhydrogel, by InvivoGen (1 mentions) Cd19 microbeads, by Miltenyi Biotec (1 mentions) Balb c mice, by Miltenyi Biotec (1 mentions) Cd4 microbeads, by Miltenyi Biotec (1 mentions)

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DuoSets (49 citations) Mouse IL-2 (13 citations) Mouse IL-2 DuoSet ELISA (4 citations) Mouse IL-2 Duoset ELISA kit (3 citations) DuoSet Mouse (2 citations)

4 protocols using mouse il 2 duoset

Quantifying IL-2 Levels in Cell Cultures

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Supernatants were collected from the cultures at 24 and 48 hrs as indicated. The levels of IL-2 in culture supernatants were then assessed by ELISA using “Mouse IL-2 Duoset” ELISA development kit from R & D Systems (Minneapolis, MN) following the assay procedure provided with manufacturer’s instructions.

Khattar M., Miyahara Y., Schroder P.M., Xie A., Chen W, & Stepkowski S.M. (2014). Interleukin-21 Is a Critical Regulator of CD4 and CD8 T Cell Survival during Priming under Interleukin-2 Deprivation Conditions. PLoS ONE, 9(1), e85882.

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Mouse aAVC-NKT Hybridoma Co-culture

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Ten thousand mouse aAVCs were co-cultured with 1 × 10⁵ NKT hybridoma (1.B2) for 24 h in 96-well round-bottom plates. Culture supernatants were analyzed for mouse IL-2 production using ELISA for cytokines with a mouse IL-2 Duo set (R&D Systems).

Yamasaki S., Shimizu K, & Fujii S.I. (2024). Tumor epitope spreading by a novel multivalent therapeutic cellular vaccine targeting cancer antigens to invariant NKT-triggered dendritic cells in situ. Frontiers in Immunology, 15, 1345037.

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T Cell Activation Assay and IL-2 ELISA

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The T cell activation assay was carried-out in triplicate in a 96 well plate using 2 × 10⁵ R8 cells irradiated at 3000 rads prior to use with 2 ×10⁵ N15 TCRαβ BW cells in each well. Concentrations of the stimulatory peptide VSV8 were added to each well ranging from 50 pg/ml to 5 μg/ml. A negative control lacked VSV8 peptide while PMA plus ionomycin was used as a positive control. The VSV8 stimulated cells and the controls were incubated for 16–18 hours overnight in a 37°C incubator, the ce ll supernatants were then harvested for an IL-2 ELISA assay.
The IL-2 ELISA assay was completed using the mouse IL-2 DuoSet and ancillary reagent kit 2 (R&D systems). T cell supernatants were diluted in media such that the O.D. 450 nm readings fall within the standard curve for the assay. The assay was then carried out following the kit instructions. Negative control values were subtracted from each sample point and concentrations in pg/ml were calculated from the standard curve. Measured IL-2 was plotted vs. the concentration of VSV8 peptide and fit to a 4-parameter logistic model.

Brazin K.N., Mallis R.J., Boeszoermenyi A., Feng Y., Yoshizawa A., Reche P.A., Kaur P., Bi K., Hussey R.E., Duke-Cohan J.S., Song L., Wagner G., Arthanari H., Lang M.J, & Reinherz E.L. (2018). The T cell antigen receptor α transmembrane domain coordinates triggering through regulation of bilayer immersion and CD3 subunit associations. Immunity, 49(5), 829-841.e6.

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Ovalbumin-Specific B and T Cell Activation

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DO11.10 and BALB/C mice were purchased from The Jackson Laboratory and housed in accordance with the policies of the IACUC of Eli Lilly and Company. BALB/C mice were primed with 10 µg/ml ovalbumin (OVA) in Alhydrogel (InvivoGen, CA, USA) at a 2:1 ratio and boosted with 10 µg/ml OVA in PBS 2 weeks after priming. BALB/C mice were sacrificed 4 days after boosting, and B cells were purified from splenocytes using CD19 microbeads (Miltenyi Biotec) for positive selection. B cells were preincubated with HM71305/LSN3359180 for 1 h before the addition of OVA protein or OVA323-339 peptide, and T cells were positively isolated from DO11.10 transgenic mice using CD4 microbeads (Miltenyi Biotec). The supernatants were collected after 18 h, and IL-2 was measured by ELISA using mouse IL-2 DuoSet (DY402, R&D Systems). FACS analysis of CD69 (H1.2F3, Biolegend), CD86 (B7-2, BD Biosciences) and H2-IA/IE (M5/114.15.3, BD Biosciences) was performed using BD LSR Fortessa, and data were analyzed by FlowJo10 (FlowJo, LLC).

Byun J.Y., Koh Y.T., Jang S.Y., Witcher J.W., Chan J.R., Pustilnik A., Daniels M.J., Kim Y.H., Suh K.H., Linnik M.D, & Lee Y.M. (2021). Target modulation and pharmacokinetics/pharmacodynamics translation of the BTK inhibitor poseltinib for model-informed phase II dose selection. Scientific Reports, 11, 18671.

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