Mouse il 2 elisa max standard

Manufactured by BioLegend

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The Mouse IL-2 ELISA MAX™ Standard is a laboratory equipment used to measure the concentration of interleukin-2 (IL-2) in mouse samples. It is a sandwich enzyme-linked immunosorbent assay (ELISA) kit that provides the necessary components to perform this specific quantitative analysis.

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Lab products found in correlation

Pe conjugated anti mouse α galcer cd1d complex l363, by BioLegend (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions)

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Mouse IL-2 ELISAs

6 protocols using mouse il 2 elisa max standard

In Vitro Generation and Characterization of Mouse Bone Marrow-Derived Dendritic Cells

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Mouse bone marrow-derived dendritic cells (BMDCs) were generated from C57BL/6 mice according to Ref. (23 (link)). At 7 day of culture, BMDCs were incubated in RPMI medium supplemented with 10% FCS, 5 µM 2-ME, 1 mM glutamine, and 1 mM sodium pyruvate for 2 h with different concentrations of free α-GalCer, fdWT bacteriophages, or fd/α-GalCer bacteriophages. The experiment OTI hybridoma cell experiment was performed by incubating BMDCs with different concentration of fdOVA, fdOVA/α-GalCer, or OVA_257–264 synthetic peptide. After the incubation, cells were washed and stained with PE-conjugated anti mouse α-GalCer:CD1d complex (L363, Biolegend) or co-cultured (50,000/well) with the mouse Vα14 iNKT hybridoma FF13 or OTI hybridoma (50,000/well) for 40 h.
PE-conjugated anti-mouse α-GalCer:CD1d complex (L363, Biolegend) antibody was used to stain DCs and fluorescence of stained cells was analyzed by FACSCanto II flow-cytometer and DIVA (Data-Interpolating Variational Analysis) software (Becton Dickinson). The IL-2 released into cell co-culture supernatants was measured by ELISA. Supernatants (0.1 ml/well) were assayed in duplicate using mouse IL-2 ELISA MAX™ Standard (Biolegend), according to the manufacturer’s instructions.

Sartorius R., D’Apice L., Barba P., Cipria D., Grauso L., Cutignano A, & De Berardinis P. (2018). Vectorized Delivery of Alpha-GalactosylCeramide and Tumor Antigen on Filamentous Bacteriophage fd Induces Protective Immunity by Enhancing Tumor-Specific T Cell Response. Frontiers in Immunology, 9, 1496.

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iNKT Cell Activation by α-GalCer Formulations

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Mouse BM-DCs (100,000/well) or human DCs (25,000/well) were incubated in RPMI-1640 medium supplemented with 10% FBS, 5 μM 2-Mercaptoethanol, 1 mM glutamine, and 1 mM sodium pyruvate for 2 h with different concentrations of free α-GalCer (1.5, 15, 150 ng/mL), fdNY-ESO-1, or fdNY-ESO-1/α-GalCer bacteriophages (0.5, 5, and 50 μg/mL, containing the above-mentioned amounts of α-GalCer, respectively). In some experiments, cells were also incubated with fdNY-ESO-1 mixed with soluble α-GalCer. After the incubation, cells were washed and co-cultured with mouse iNKT cells (50,000/well) for 40 h. IL-2 released into co-culture supernatants was measured via ELISA. Supernatants (0.1 mL/well) were assayed in duplicate using mouse IL-2 ELISA MAX™ Standard (Biolegend) according to the manufacturer’s instructions.

Manco R., D’Apice L., Trovato M., Lione L., Salvatori E., Pinto E., Compagnone M., Aurisicchio L., De Berardinis P, & Sartorius R. (2023). Co-Delivery of the Human NY-ESO-1 Tumor-Associated Antigen and Alpha-GalactosylCeramide by Filamentous Bacteriophages Strongly Enhances the Expansion of Tumor-Specific CD8+ T Cells. Viruses, 15(3), 672.

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Phage-Mediated Antigen Cross-Presentation

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fdOVA encapsulated directly or by the post encapsulation method was released from microparticles, and 1 × 10⁶/mL BM-DCs were incubated overnight with 0.06 μg/mL of released bacteriophages (Fig. 6) [10 (link)].
Alternatively, BM-DCs (1 × 10⁶/mL) were left to adhere to multiwell plates. Phage-loaded MPs were resuspended in PBS, and a volume of MPs containing 0.06 μg/mL of encapsulated fdOVA was immediately added to adherent BM-DCs.
BM-DCs were then washed twice and plated at 100,000 cells/well, and the antigen cross presentation was detected by adding the OTI hybridoma cell line B3Z (50,000 cells/well) to the culture. The IL-2 released in the culture medium by activated B3Z cells was measured after 40 h using the supernatants of the co-cultures (0.1 mL/well) and a mouse IL-2 ELISA MAX™ Standard (Biolegend).

Jamaledin R., Sartorius R., Di Natale C., Onesto V., Manco R., Mollo V., Vecchione R., De Berardinis P, & Netti P.A. (2023). PLGA microparticle formulations for tunable delivery of a nano-engineered filamentous bacteriophage-based vaccine: in vitro and in silico-supported approach. Journal of Nanostructure in Chemistry.

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Quantifying OVA257-264 Presentation by DCs

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1 × 10⁶/mL immature BM-DCs were incubated for 3 h with different concentrations of OVA_257–264 synthetic peptide (from 0.0004 to 4 µM) in presence of 1 µg/mL of LPS, MPLA, or PGDG 2. As control, DCs incubated only with medium were used. After the incubation, cells were washed and co-cultured (50,000/well) with the OTI hybridoma cell line B3Z (50,000/well) for 40 h. The IL-2 released into cell co-culture supernatants was measured by ELISA. Supernatants (0.1 mL/well) were assayed in duplicate using mouse IL-2 ELISA MAX™ Standard (Biolegend, San Diego, CA, USA), according to the manufacturer’s instructions. B3Z OTI hybridoma cell line, recognizing the OVA_257–264 SIINFEKL determinant, was growth in complete RPMI 1640 (10% FCS 100 U/mL penicillin, 100 μg/mL streptomycin 1% Glutamine, 1% NEM, 1% Sodium Pyruvate, 50 µM 2-Mercaptoethanol) and was a kind gift of Dr. Bénédicte Manoury (INEM, INSERM U1151-CNRS UMR 8253, Hôpital Necker, Paris, France).

Manzo E., Gallo C., Sartorius R., Nuzzo G., Sardo A., De Berardinis P., Fontana A, & Cutignano A. (2019). Immunostimulatory Phosphatidylmonogalactosyldiacylglycerols (PGDG) from the Marine Diatom Thalassiosira weissflogii: Inspiration for a Novel Synthetic Toll-Like Receptor 4 Agonist. Marine Drugs, 17(2), 103.

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Evaluating MHC I-Presented OVA Epitope

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1 × 10⁶/mL BM-DCs were incubated overnight with different concentrations (from 0.6 to 6 ug/mL) of free fdOVA filamentous bacteriophage or fdOVA released from MPs. As a control, BM-DCs incubated only with the medium were used. After the incubation, cells were washed twice to remove excess bacteriophages, then co-cultured (100,000/well) with the OTI hybridoma cell line B3Z (50,000/well) for 40 h. B3Z OTI hybridoma cell line, recognizing the OVA (257–264) SIINFEKL determinant, was grown in complete RPMI1640 (10% FCS, 100 U/mL penicillin, 100 μg/mL streptomycin 1% Glutamine, 1% NEM, 1% Sodium Pyruvate, 50 µM 2-Mercaptoethanol). Recognition of the major histocompatibility complex (MHC I)-presented OVA peptide 257–264 (SIINFEKL) by B3Z T cell receptor led to the transcriptional activation of the IL-2 promoter element, resulting in production of IL-2, which correlates with the uptake and processing of the fdOVA and the presentation of OVA (257–264) peptide in MHC I. The amount of IL-2 released into cell co-culture supernatants was measured by ELISA. Supernatants of co-cultures (0.1 mL/well) were assayed in duplicate using mouse IL-2 ELISA MAX Standard (Biolegend, San Diego, CA), according to the manufacturer’s instructions (Figure 3).

Jamaledin R., Sartorius R., Di Natale C., Vecchione R., De Berardinis P, & Netti P.A. (2020). Recombinant Filamentous Bacteriophages Encapsulated in Biodegradable Polymeric Microparticles for Stimulation of Innate and Adaptive Immune Responses. Microorganisms, 8(5), 650.

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Quantifying Murine IL-2 Production

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On the supernatant of the OT-1 cells activated by the BMDC, the production of murine IL-2 was measured by ELISA. IL-2 was quantified using the Mouse IL-2 ELISA MAX™ Standard from Biolegend. All ELISA procedures were performed according to the manufacturer's protocol.

Duvallet E., Boulpicante M., Yamazaki T., Daskalogianni C., Prado Martins R., Baconnais S., Manoury B., Fahraeus R, & Apcher S. (2016). Exosome-driven transfer of tumor-associated Pioneer Translation Products (TA-PTPs) for the MHC class I cross-presentation pathway. Oncoimmunology, 5(9), e1198865.

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