Recombinant human ifn γ

Manufactured by ProSpec

Recombinant human IFN-γ is a laboratory-produced protein that is structurally and functionally similar to the naturally occurring human interferon gamma. It is a cytokine that plays a key role in the immune response.

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Lab products found in correlation

Lsr fortessa 4, by BD (1 mentions) Recombinant human ifn α2, by BioLegend (1 mentions) Bortezomib, by Merck Group (1 mentions)

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IFN-γ (44 citations)

2 protocols using recombinant human ifn γ

Phosphorylation of STAT Proteins in Whole Blood

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200 μl of whole blood was stained with anti-human CD4-BV510, anti-human CD8-APC-H7 and anti-human CD38-APC at previously determined optimum dilution for 15 mins at 37°C with or without recombinant human IFN-α2 (Biolegend) or recombinant human IFN-γ (ProSpec) at 100 ng/mL. Whole blood was lysed and fixed with Phosflow Lyse/fix 1x solution (BD Biosciences) and lymphocytes permeabilized with BD Perm III solution (BD Biosciences) according to the manufacturer’s instructions. Cells were then resuspended in 50 μl of staining buffer containing anti-human pSTAT1-PECF594, anti-human pSTAT4-AF488 and anti-human pSTAT5-PE at previously determined optimum dilution for one hour at room temperature. Cells were washed and resuspended in staining buffer. Samples were acquired using a LSR Fortessa 4 (BD Biosciences) with Diva software, and analyzed using FlowJo software (version 6.0).

Burel J.G., Apte S.H., Groves P.L., Klein K., McCarthy J.S, & Doolan D.L. (2016). Reduced Plasmodium Parasite Burden Associates with CD38+ CD4+ T Cells Displaying Cytolytic Potential and Impaired IFN-γ Production. PLoS Pathogens, 12(9), e1005839.

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Fluorescence-Based Immune Cell Profiling

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The following antibodies were used: CD14-FITC (61D3), CD16-BV510 (3G8), SLAMF7-PE (162), CD3-PE-Cy7 (OKT3), CD19-PE-Cy7 (SJ25C1), CD57-PE-Cy7 (TB01), CD66b-APC (G10F5), CXCL10-PerCp-eFluor710 (4NY8UN), EAT-2-APC (LS-C240730), YY1-Alexa647 (H-10), Blimp-1-DyLight650 (3H2-E8), CCR5-PerCp-eFluor710 (NP-6G4) and SLAMF7 (162.1) (used for cross-linking). The SLAMF7-Fc recombinant protein was designed and produced similar to mCRACC-Fc as previously described (25 (link)), with the following modifications: a human SLAMF7 extracellular domain was swapped for the murine SLAMF7 extracellular domain, the murine IgG Fc portion was switched to a human IgG4 domain to reduce Fc receptor binding and ADCC, and S228P and L235E mutations were made in the IgG4 domain to further reduce interactions with Fc receptors (31 (link)). Recombinant universal IFN⍺ (PBL Assay Bioscience) and Recombinant human IFNγ (ProSpec) were used at 100 IU/mL unless otherwise noted. SHP1/2 inhibitor (NSC87877) (Millipore Sigma) was used at 10μM. SHIP1 inhibitor (3AC) (Millipore Sigma) was used at 5μM. CD45 inhibitor (CAS 345630–40-2) (Millipore Sigma) was used at 1μM. Bortezomib (EMD Millipore) was used at 100nM. All HDACi’s (Selleck chemicals) were used at concentrations indicated in relevant figure legends.

O’Connell P., Pepelyayeva Y., Blake M.K., Hyslop S., Crawford R.B., Rizzo M.D., Pereira-Hicks C., Godbehere S., Dale L., Gulick P., Kaminski N.E., Amalfitano A, & Aldhamen Y.A. (2018). SLAMF7 is a critical negative regulator of interferon-⍺-mediated CXCL10 production in chronic HIV infection. Journal of immunology (Baltimore, Md. : 1950), 202(1), 228-238.

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