Recombinant mouse ifn γ and il 4

Manufactured by R&D Systems

Recombinant mouse IFN-γ and IL-4 are cytokine proteins produced using recombinant DNA technology. IFN-γ is a type II interferon, and IL-4 is a type I cytokine. These proteins are used for research purposes.

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Lab products found in correlation

Anti il4 blocking antibody clone 11b11, by Thermo Fisher Scientific (2 mentions) Anti h3k27ac ab4729 antibody, by Abcam (2 mentions) Phospo stat1, by Merck Group (2 mentions) Stat6, by Merck Group (2 mentions) Phospho stat6, by Merck Group (2 mentions) Phospho stat1 alexafluor 647, by BD (2 mentions) Phospho stat6 pe, by BD (2 mentions) Alexa fluor 488 goat anti rabbit secondary antibody, by Thermo Fisher Scientific (2 mentions) Stat1, by Merck Group (2 mentions) Phospho erk1 2, by Merck Group (2 mentions) Phospho akt, by Merck Group (2 mentions) Anti actin a4700, by Merck Group (2 mentions) Horseradish peroxidase conjugated streptavidin, by Merck Group (1 mentions) Microtainer serum separator tube, by BD (1 mentions) Hrp conjugated antibody, by Southern Biotech (1 mentions) Complete freund adjuvant (cfa), by Merck Group (1 mentions) Ovalbumin (ova), by Merck Group (1 mentions)

3 protocols using recombinant mouse ifn γ and il 4

Signaling Mechanisms in Macrophage Activation

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Recombinant mouse IFN-γ and IL-4 were from R&D Systems. Unless otherwise stated, cytokines were used at a concentration of 100ng/ml and 10ng/ml, respectively. Anti-IL4 blocking antibody (clone 11B11) was from eBioscience. CD11B (clone M1/70), F4/80 (BM8), NOS2 (CXNFT) antibodies for flow cytometry were from eBioscience. Antibodies used in ChIP experiments against Stat1 (sc-592), Stat6 (sc-981), MYC (sc-764), JUNB (sc-46x), C/EBPβ (sc-150X) were all from Santa Cruz. The anti-H3K27Ac (ab4729) antibody was from Abcam. The anti-Pu.1 rabbit polyclonal antibody was generated in-house against the N-terminus of Pu.1 (aa. 1-100; NP_035485.1) 32 (link). Antibodies used in western blot experiments: Stat1 (#9172), phospo-Stat1 (Tyr701, #9171), Stat6 (#9362), phospho-Stat6 (Tyr641, #9361), Erk1/2 (#9102), phospho-Erk1/2 (Thr202/Tyr204, #9101), phospho-Akt (Ser473, #9271); anti-actin (A4700) from Sigma-Aldrich. The following antibodies were used for flow cytometry: CD11b (clone M1/70), F4/80 (BM8), Nos2 (CXNFT) from eBioscience; phospho-Stat1-AlexaFluor 647 (Cat. 612597) and phospho-Stat6-PE (Cat. 558252) from BD Biosciences. The following antibodies were used for immunofluorescence: Alexafluor 488 goat anti-rabbit secondary antibody (Cat. R37116) and DAPI (Cat. 62247) from Thermo Scientific; phospho-Stat1-AlexaFluor 647 (Cat. 612597) and phospho-Stat6-PE (Cat. 558252) from BD Biosciences.

Piccolo V., Curina A., Genua M., Ghisletti S., Simonatto M., Sabò A., Amati B., Ostuni R, & Natoli G. (2017). Opposing macrophage polarization programs show extensive epigenomic and transcriptional cross talks. Nature immunology, 18(5), 530-540.

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OVA-Specific Antibody and Cytokine Responses

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Mice were immunized subcutaneously with OVA (Sigma) in CFA (Sigma) and euthanized after 14 days. Blood was collected and allowed to coagulate in the refrigerator in Microtainer Serum Separator tube (BD). Serum was isolated by centrifugation. For ELISA to detect OVA-specific Ig, plates were coated with 2 mg ml⁻¹ OVA. Antibody levels in serially diluted serum samples were analysed with HRP-conjugated antibodies (1:4,000) specific for mouse IgM (1021-05), IgG (1030-05), IgG1 (1070-05), and IgG2c (1079-05) (SouthernBiotech) following manufacture's recommended protocols. For ex vivo cytokine production, splenocytes were isolated 14 days following OVA-CFA immunization and cultured in the presence of OVA (100 μg ml⁻¹). ELISAs were performed with purified anti-IFNγ (Biolegend 505707) and anti-IL-4 mAb (Biolegend 504102) as capture antibodies, the corresponding biotinylated anti-IFNγ (Biolegend 505804, 1 μg ml⁻¹)and anti-IL-4 mAb (BD Biosciences 554390 1 μg ml⁻¹), horseradish peroxidase-conjugated streptavidin (Sigma), and the TMB microwell peroxidase substrate and stop solution (Kirkegaard & Perry Labs, Inc.) according to the recommended protocol (Biolegend). Recombinant mouse IFNγ and IL-4 (R&D Systems) was used as standards.

Thornton T.M., Delgado P., Chen L., Salas B., Krementsov D., Fernandez M., Vernia S., Davis R.J., Heimann R., Teuscher C., Krangel M.S., Ramiro A.R, & Rincón M. (2016). Inactivation of nuclear GSK3β by Ser389 phosphorylation promotes lymphocyte fitness during DNA double-strand break response. Nature Communications, 7, 10553.

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Signaling Mechanisms in Macrophage Activation

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