Anti il22 percp efluor710

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Anti-IL22 PerCP-Efluor710 is a monoclonal antibody labeled with PerCP-eFluor 710 fluorescent dye. It is designed for the detection and analysis of interleukin-22 (IL-22) expression in cells using flow cytometry.

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4 protocols using anti il22 percp efluor710

Multiparameter Flow Cytometry of Immune Cells

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Cryopreserved cells were thawed, washed, and permeabilized with Perm/Wash solution (BD Biosciences). Cells were then incubated at 4°C for 1 h with fluorescence-conjugated antibodies directed against surface antigens and intracellular cytokines. For detection of the transcription factor FOXP3, a nuclear permeabilization buffer (eBioscience, San Diego, CA, USA) was used in place of the Perm/Wash solution as per manufacturer’s protocol. The following fluorescence-conjugated antibodies were used: anti-CD3 PacBlue, anti-γδ TCR PE, anti-CD27 PECy7, anti-CD25 APC, anti-IFNγ Alexafluor 700, anti-IL17A Alexafluor 647, anti-Ki67FITC (BD biosciences, San Jose, CA, USA); anti-CD45RA QDot655, anti-CD8 QDot605, anti-CD4 QDot605 (Invitrogen, Eugene, OR, USA); and anti-CD39 FITC, anti-IL22 PerCP-Efluor710, and anti-FOXP3 PE (eBioscience, SanDiego, CA, USA).
The entire sample was acquired on a BD LSRFortessa Flow Cytometer using FACSDiva software (BD Biosciences, San Jose, CA, USA). Compensation was performed using compensation beads.

Whittaker E., Nicol M., Zar H.J, & Kampmann B. (2017). Regulatory T Cells and Pro-inflammatory Responses Predominate in Children with Tuberculosis. Frontiers in Immunology, 8, 448.

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Multiparameter Flow Cytometry of CD8+ T Cell Subsets

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To analyze the frequency of CD8+ T cell subsets, the harvested cells were washed once with PBS and evaluated by flow cytometry. In brief, the 10⁶ cells/100ul staining buffer (PBS + 2%FCS) were first stained with Amcyan-conjugated anti-human CD8 monoclonal antibodies (BD Biosciences, US) for 30 minutes in the dark at 4°c. After surface staining, the cells were fixed and permeabilized using LEUCOPERM^™ (BIO-RAD, BUF09B, US) according to the manufacturer’s instructions. The cells were stained with anti-IFN-γ-FITC, anti-TNF-α-PE-Cy^™7, anti-IL-10-eFluor^®450, anti-IL-17-PE, anti-IL-21-eFluor^®660, anti-IL-22-PerCP-eFluor^®710 (eBioscience, US) and anti- IL-4-APC-Cy^™7 (BioLegend, US).

Salehi Z., Doosti R., Beheshti M., Janzamin E., Sahraian M.A, & Izad M. (2016). Differential Frequency of CD8+ T Cell Subsets in Multiple Sclerosis Patients with Various Clinical Patterns. PLoS ONE, 11(7), e0159565.

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Multiparameter Flow Cytometric Analysis of Immune Cell Subsets

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Cryopreserved cells were thawed, washed, and permeabilized with Perm/wash solution (BD Biosciences). Cells were then incubated at 4 °C for 1 hour with fluorescence-conjugated antibodies directed against surface antigens and intracellular cytokines. For detection of the transcription factor FOXP3, a nuclear permeabilisation buffer (eBioscience) was used in place of the Perm/wash solution as per manufacturer’s protocol. The following fluorescence-conjugated antibodies were used: anti-CD3 PacBlue, anti-GDTCR PE, anti-CD27 PECy7, anti-CD25 APC, anti-IFNγ Alexafluor 700, anti-IL17A Alexafluor 647, anti-Ki67FITC (all BD biosciences, San Jose, CA); anti-CD45RA QDot655, anti-CD8 QDot605, anti-CD4 QDot605 (all Invitrogen, Eugene, Or); anti-CD39 FITC, anti-IL22 PerCP-Efluor710, anti-FOXP3 PE (all eBioscience, SanDiego, CA).
The entire sample was acquired on a BD LSR Fortessa Flow Cytometer (model 649225B7, power 690 W, manufactured Sept 2010) configured with 4 lasers and 22 detectors using FACSDiva software (BD Bioscience, San Jose, CA, USA). Compensation for overlap of fluorescence detection was performed using compensation beads. Multiparameter panel development included evaluation of appropriate staining controls of antibody and fluorochrome interactions and of spectral overlap using control blood samples.

Whittaker E., Nicol M.P., Zar H.J., Tena-Coki N.G, & Kampmann B. (2018). Age-related waning of immune responses to BCG in healthy children supports the need for a booster dose of BCG in TB endemic countries. Scientific Reports, 8, 15309.

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Detailed Phenotyping of Immune Cells

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Anti-CD14 and anti-CD19 APC-H7 (both at 1:100), anti-CD25 PECy7 (1:50), anti-CD62L V450 (1:50), anti-CD127 PECy7 (1:100), anti-CD161 FITC (1:10), anti-IFNγ V450 (1:200), anti-Ki67 FITC (1:10) were from BD Biosciences. Anti-CD3 ECD (1:100) and anti-CD8β PE (1:50) were from Beckman Coulter. Anti-Vα7.2 FITC (1:10) and PE (1:20), anti-CD45 Alexa Fluor 700 (1:400), anti-CD45RO PECy7 (1:50), anti-CCR9 Alexa Fluor 647 (1:20), anti-Ki67 Brilliant Violet 421 (1:20), anti-CCR7 Brilliant Violet 421 (1:20) were from BioLegend. Anti-CD161 PerCP-Cy5.5 (1:20), anti-IL-22 PerCP-eFluor 710 (1:50) were from eBioscience. Anti-IL-18R PE (1:20), and anti-PLZF APC (1:10) were from R&D systems. Anti-CD4 Qdot 655 (1:400), anti-CD8α Qdot 605 (1:800), anti-CD45 Qdot 705 (1:100), live/dead aqua (1:100) and near infrared fixable (1:400) cell stain were from Invitrogen. Cell surface staining was performed using directly conjugated antibodies and fixed in Cytofix/Cytoperm (BD Biosciences). Intracellular staining was performed using the appropriate mAb’s in Perm/Wash (BD Biosciences). Samples were acquired on an LSRII flow cytometer (BD Biosciences) equipped with 405, 488, 532 and 647 nm lasers. Single-stained polystyrene beads (BD Biosciences) were used for compensation purposes. Software-based compensation was performed using the compensation platform in FlowJo software version 9.5 (Tree Star).

Leeansyah E., Loh L., Nixon D.F, & Sandberg J.K. (2014). Acquisition of innate-like microbial reactivity in mucosal tissues during human fetal MAIT-cell development. Nature Communications, 5, 3143.

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