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Anti ifn γ pe cy7 xmg1.2

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The Anti–IFN-γ PE-Cy7 (XMG1.2) is a fluorochrome-conjugated monoclonal antibody that binds to the interferon-gamma (IFN-γ) protein. It is commonly used in flow cytometry applications to detect and quantify IFN-γ-producing cells.

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Lab products found in correlation

Brefeldin a, by Merck Group (4 mentions) Live dead fixable blue dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Anti il 2 jes 5h4 apc cy7, by BD (2 mentions) Anti il 10 jes5 16e3 apc, by Thermo Fisher Scientific (2 mentions) Anti il17a ebi17b7 fitc, by Thermo Fisher Scientific (2 mentions) C maf percp efluor symof1 710, by Thermo Fisher Scientific (2 mentions) Igg2bk isotype bmg2b efluor 710 control, by Thermo Fisher Scientific (2 mentions) Cytofix cytoperm kit, by BD (2 mentions) Anti cd4 allophycocyanin clone rm4 5, by BD (2 mentions) Anti tnf α pe mp6 xt22, by BD (2 mentions) Il 2 allophycocyanin cy7 jes6 5h4, by BD (2 mentions) Il 17a percp cy5.5 i7b7, by Thermo Fisher Scientific (2 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Streptavidin bv421, by BioLegend (1 mentions) Fitc anti cd11b m1 70, by BioLegend (1 mentions) Facscanto 2 device, by BD (1 mentions) Fcsexpress 5flow software, by De Novo Software (1 mentions) Anti cd25 fitc 7d4, by BD (1 mentions) Percp 7 aad, by BioLegend (1 mentions)

Close spelling variants of this product

IFN-γ (389 citations) Anti-IFN-γ (150 citations) Anti-IFN-γ-PE (76 citations) IFN-γ-PE (64 citations) IFN-γ-PE-Cy7 (42 citations) IFN-γ (XMG1.2) (35 citations) Anti-IFN-γ (XMG1.2) (27 citations) Anti-IFN-γ PE-Cy7 (25 citations)

5 protocols using anti ifn γ pe cy7 xmg1.2

1

T Cell Stimulation and Cytokine Analysis

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After differentiation, T cells were re-stimulated with anti-CD3 and anti-CD28 at 2 μg/ml each or PDBU (Sigma-Aldrich) and Ionomycin (Calbiochem) at 500 ng/ml each (TH17) for 4 hours, with Brefeldin A (10 μg/ml, Sigma-Aldrich) added for the last 2 and 4 hours of culture, respectively. Surface markers were stained in PBS together with LIVE/DEAD Fixable Blue Dead Cell Stain Kit (Molecular Probes). For intracellular cytokine detection, cells were fixed with 2% formaldehyde, permeabilized with permeabilization buffer (eBioscience) and stained with: anti-IL-2 (JES-5H4) APC-Cy7 (BD), anti-IFN-γ (XMG1.2) PE-Cy7 (BD) and anti-IL-4 (11B11) PE, anti-IL-10 (JES5-16E3) APC, anti-IL17A (eBi17B7) FITC (eBioscience). For Foxp3 and c-Maf staining, cells were fixed with Fixation/permeabilization buffers and stained with Foxp3 (FJK-16S) PE or c-Maf PerCP-eFluor (symOF1) 710 with IgG2bK isotype (BMG2b) eFluor 710 control (eBioscience).
Gabryšová L., Alvarez-Martinez M., Luisier R., Cox L.S., Sodenkamp J., Hosking C., Pérez-Mazliah D., Whicher C., Kannan Y., Potempa K., Wu X., Bhaw L., Wende H., Sieweke M.H., Elgar G., Wilson M., Briscoe J., Metzis V., Langhorne J., Luscombe N.M, & O’Garra A. (2018). c-Maf controls immune responses by regulating disease-specific gene networks and repressing IL-2 in CD4+ T cells. Nature immunology, 19(5), 497-507.
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2

T Cell Stimulation and Cytokine Analysis

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After differentiation, T cells were re-stimulated with anti-CD3 and anti-CD28 at 2 μg/ml each or PDBU (Sigma-Aldrich) and Ionomycin (Calbiochem) at 500 ng/ml each (TH17) for 4 hours, with Brefeldin A (10 μg/ml, Sigma-Aldrich) added for the last 2 and 4 hours of culture, respectively. Surface markers were stained in PBS together with LIVE/DEAD Fixable Blue Dead Cell Stain Kit (Molecular Probes). For intracellular cytokine detection, cells were fixed with 2% formaldehyde, permeabilized with permeabilization buffer (eBioscience) and stained with: anti-IL-2 (JES-5H4) APC-Cy7 (BD), anti-IFN-γ (XMG1.2) PE-Cy7 (BD) and anti-IL-4 (11B11) PE, anti-IL-10 (JES5-16E3) APC, anti-IL17A (eBi17B7) FITC (eBioscience). For Foxp3 and c-Maf staining, cells were fixed with Fixation/permeabilization buffers and stained with Foxp3 (FJK-16S) PE or c-Maf PerCP-eFluor (symOF1) 710 with IgG2bK isotype (BMG2b) eFluor 710 control (eBioscience).
Gabryšová L., Alvarez-Martinez M., Luisier R., Cox L.S., Sodenkamp J., Hosking C., Pérez-Mazliah D., Whicher C., Kannan Y., Potempa K., Wu X., Bhaw L., Wende H., Sieweke M.H., Elgar G., Wilson M., Briscoe J., Metzis V., Langhorne J., Luscombe N.M, & O’Garra A. (2018). c-Maf controls immune responses by regulating disease-specific gene networks and repressing IL-2 in CD4+ T cells. Nature immunology, 19(5), 497-507.
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3

Quantifying Antigen-Specific T-Cell Responses

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A total of 1–2×106 lung MNCs were stimulated in vitro in V-bottom 96-well plates at 37°C in media containing anti-CD49d (1 mg/ml) and anti-CD28 (1 mg/ml) without antigen or in the presence of protein or peptide antigen (5 μg/ml) for 1 h, plus 6 h in the presence of 10 mg/ml brefeldin A (Sigma-Aldrich), after which cells were maintained at 4°C for 2–8h before staining.
Cells were stained for surface markers using anti–CD4-allophycocyanin (clone RM4-5; BD Pharmingen) before fixation and permeabilization using Cytoperm/Cytofix kit (BD Pharmingen) as per manufacturer’s instructions, and subsequently stained for intracellular cytokines with anti–IFN-γ PE-Cy7 (XMG1.2; BD Pharmingen), anti–TNF-α–PE (MP6-XT22; BD Pharmingen), IL-2–allophycocyanin–Cy7(JES6-5H4; BD Pharmingen), and IL-17A–PerCP–Cy5.5 (I7B7; eBiosciences).
Antigen-specific cell numbers determined from Nucleocounter (ChemTec, Deerfield Beach. FL, USA) enumeration in combination with ICS data of % cytokine-producing cells after stimulation. Non-specific background cytokine values were determined for each combinatorial Boolean gate and subtracted.
Woodworth J.S., Cohen S.B., Moguche A.O., Plumlee C.R., Agger E.M., Urdahl K.B, & Andersen P. (2016). Subunit vaccine H56/CAF01 induces a population of circulating CD4 T cells that traffic into the Mycobacterium tuberculosis-infected lung. Mucosal immunology, 10(2), 555-564.
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4

Multidimensional Immunophenotyping of Splenic Cells

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Splenic single-cell suspensions were incubated with fluorochrome-conjugated Abs (FITC, PE, PECy7, PerCPCy5.5, allophycocyanin, APCCy7) in PBS for 30 min at 4 °C. For intracellular staining, the eBioscience Foxp3/Transcription Factor Staining Buffer Set was used, according to the manufacturer’s instructions. Stained cells were analyzed using FACSCanto II devices (BD Biosciences) and FCS Express5Flow software (DeNovo). Viable lymphocytes were gated based on forward scatter and side scatter prior to further gating of fluorescent-labeled populations. The following Abs were used: anti-CD4–APC/Fire™750-RM4-5, anti-CD3-BV421 17A2, anti-B220–APC RA3-6B2 7D4, anti-CD44-PerCPCy5.5 IM7, anti-CD11b–FITC M1/70, anti-IL-17-AF614 TC11-18H10.1, anti-CD19- PerCPCy5.5 6D5, anti-CD25–APCCy7 PC61, anti-mouse Ki-67-AF488 16A8, viability staining solution-PerCP 7-AAD, anti-PD-1-APC RMP1-30, anti-CD244-FITC m2B$(B6)458.1, anti-LAG-3-PE C9B7W, anti-TIM-3-PerCP B8.2C12, anti-CTLA-4_PECy7 UC104B9, anti-CXCR5-bio L138D7, Streptavidin-BV421, anti-Bcl-6-PA IG191E/A8 (all from Biolegend). Anti-CD8a–PerCP 53-6.7, anti-CD25–FITC 7D4, anti-CD62L-APC MEL-14, anti-IFNγ-PECy7 XMG1.2, anti-IL-4- PE 11B11 (RUO) (all from BD Biosciences) and anti-Foxp3–PE FJK-16s (all from eBioscience).
Chemnitz J., Pieper D., Stich L., Schumacher U., Balabanov S., Spohn M., Grundhoff A., Steinkasserer A., Hauber J, & Zinser E. (2019). The acidic protein rich in leucines Anp32b is an immunomodulator of inflammation in mice. Scientific Reports, 9, 4853.
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5

Quantifying Antigen-Specific T-Cell Responses

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A total of 1–2×106 lung MNCs were stimulated in vitro in V-bottom 96-well plates at 37°C in media containing anti-CD49d (1 mg/ml) and anti-CD28 (1 mg/ml) without antigen or in the presence of protein or peptide antigen (5 μg/ml) for 1 h, plus 6 h in the presence of 10 mg/ml brefeldin A (Sigma-Aldrich), after which cells were maintained at 4°C for 2–8h before staining.
Cells were stained for surface markers using anti–CD4-allophycocyanin (clone RM4-5; BD Pharmingen) before fixation and permeabilization using Cytoperm/Cytofix kit (BD Pharmingen) as per manufacturer’s instructions, and subsequently stained for intracellular cytokines with anti–IFN-γ PE-Cy7 (XMG1.2; BD Pharmingen), anti–TNF-α–PE (MP6-XT22; BD Pharmingen), IL-2–allophycocyanin–Cy7(JES6-5H4; BD Pharmingen), and IL-17A–PerCP–Cy5.5 (I7B7; eBiosciences).
Antigen-specific cell numbers determined from Nucleocounter (ChemTec, Deerfield Beach. FL, USA) enumeration in combination with ICS data of % cytokine-producing cells after stimulation. Non-specific background cytokine values were determined for each combinatorial Boolean gate and subtracted.
Woodworth J.S., Cohen S.B., Moguche A.O., Plumlee C.R., Agger E.M., Urdahl K.B, & Andersen P. (2016). Subunit vaccine H56/CAF01 induces a population of circulating CD4 T cells that traffic into the Mycobacterium tuberculosis-infected lung. Mucosal immunology, 10(2), 555-564.
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