The following mAbs were used: anti-CD20 BUV395, anti-CD10 APC, anti-CD4 BUV737, anti-CD4 APCCy7, anti-CD25 PECy7, anti-CD27 PECy7, anti-CD27 PE, anti-CD45RA BV605, anti-CXCR5 A647, anti-IgG APC, anti-IgG PE, anti-IgG BV605, anti-CD8 BUV395, anti-PD1 BV605, Streptavidin-PerCPCy5.5, anti-IgM PerCPCy5.5, anti-CD3 BV421, anti-IL-2 BV711, anti-IL-9 PerCPCy5.5, anti-IL-13 BV421, anti-IL-17F BV786, anti-IFN-γ BV605, anti-TNF-α BUV395, anti-CD19 BV711, anti-CD34 FITC, anti-CCR6 PE (all from Becton Dickinson); anti-CCR7 PECy7, anti-CD127 BV650, anti-IL-17A APCCy7, anti-CD20 Pacific Blue, anti-FoxP3 PE, anti-CXCR3 BV421 (BioLegend); anti-CD45RA PerCPCy5.5 (eBioscience); anti-CCR7 FITC (R&D Systems); anti-IgA-biotin (SouthernBiotech); anti-IL-4 PECy7, anti-IL-21 e660, anti-IL-22 PE (Thermo Fisher Scientific).
Anti il 22 pe
Manufactured by Thermo Fisher Scientific
Anti-IL-22 PE is a fluorescent-labeled antibody used for the detection and quantification of interleukin-22 (IL-22) in various laboratory applications. It is a tool for researchers studying immune responses and cytokine signaling.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd4 apc cy7, by BD (2 mentions)
Anti cd3 bv421, by BD (2 mentions)
Anti cd34 fitc, by BD (2 mentions)
Anti cd25 pe cy7, by BD (2 mentions)
Anti ccr6 pe, by BD (2 mentions)
Anti cd27 pe, by BD (2 mentions)
Apc anti cd10, by BD (2 mentions)
Anti ccr7 fitc, by R&D Systems (2 mentions)
Anti igg apc, by BD (2 mentions)
Anti cd27 pe cy7, by BD (2 mentions)
Anti il 2 bv711, by BD (2 mentions)
Anti il 17f bv786, by BD (2 mentions)
Anti tnf α buv395, by BD (2 mentions)
Anti il 13 bv421, by BD (2 mentions)
Anti il 17a apccy7, by BioLegend (2 mentions)
Anti ifn γ bv605, by BD (2 mentions)
Anti cd19 bv711, by BD (2 mentions)
Anti cd127 bv650, by BioLegend (2 mentions)
Anti cd20 pacific blue, by BioLegend (2 mentions)
Anti cxcr3 bv421, by BioLegend (2 mentions)
6 protocols using anti il 22 pe
1
Comprehensive Immune Cell Profiling
2
Multiparametric Flow Cytometry of PBMC Subsets
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Cryopreserved PBMCs and their subpopulations were analysed with a 28-color flow cytometry panel, as previously described [34 (link)]. The following mAbs were used: anti-CD20 BUV805, anti-CD10 APC, anti-Vαβ TCR BUV737, anti-CD4 APCCy7, anti-CD25 PECy7, anti-CD27 PECy7, anti-CD27 PE, anti-CD45RA PerCpCy5, anti- CXCR5 BUV615, anti-IgG APC, anti IgG BB660, anti-IgD BV480, anti-IgG BV605, anti-IgA1/A2 PECy5, anti-CD8 BUV496, anti-CD21 BUV563, anti-PD1 BV605, anti-IgM PerCPCy5.5, anti-IgM APC R700, anti-CD3 BV421, anti-IL-2 BV711, anti-IL-9 PerCPCy5.5, anti-IL-13 BV421, anti-IL-17F BV786, anti-IFN-γ BV605, anti-TNF-α BUV395, anti-CD19 BV711, anti-CD34 FITC, anti-CCR6 PE, anti-CD45RA BUV395, anti-CXCR5 BV615 (all from Becton Dickinson); anti-CD20 Pacific Blue, anti-CCR7 PECy7, anti-CD127 BV650, anti-IL-17A APCCy7, anti-CD20 BUV805, anti-CXCR3 BV421, anti-CD3 BV570 (BioLegend); (OptiBuild); anti-CCR7 FITC (R&D Systems); anti-IL-4 PECy7, anti-IL-21 e660, anti-IL-22 PE (Thermo Fisher Scientific).
3
Characterizing Gut Immune Responses in Mice
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Strains (5 × 108 CFU/day/mice) were administered by intragastric gavage to WT conventional BALB/c mice for 5 days. Colon samples were removed at sacrifice and stored in RNAlater® storage solution (Ambion, Life Technologies, Foster City, CA, USA) at − 80 °C until qRT-PCR analysis. MLN and intestine were harvested and immediately processed for flow cytometry. Cell suspensions of MLN and intestine (3–5 × 106 cells, in RPMI1640 supplemented by 10% FCS, 2 mM l -glutamine, 2 mM HEPES, 40 mg/ml gentamycine) were stimulated using the Leukocyte Activation Cocktail containing BD GolgiPlug (BD Biosciences) (1 μl/ml of cell suspension) for 5 h. Cells were stained by mAbs anti-mouse CD11c eFluor450, CD11b eFluor 450, B220 eFluor 450, CD3 eFluor 450, CD117 Alexa Fluor 700, NK1.1 PerCP-Cy5.5, NKp46 FITC (provided by eBioscience), CD4 APC-H7, CD90.2 BV 500; CD45RB BV 605, MHCII BV 650 (provided by BD Bioscience, san Jose, CA, USA). Subsequently, cells were permeabilized and fixed using the Transcription Factor Buffer Set (BD Bioscience), and intracellular staining was performed using mAbs anti-IL-17A eFluor450, anti-FOXP3 PE-Cy7, anti-IL-22 PE and RORgt APC (eBioscience). Flow cytometry data were analyzed using software FlowJo. Gating strategy and representative dot plots for control and BIO5768 treated mice are presented in Figs. S1 and S2 .
4
Profiling Rat Liver Lymphocytes
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Rat liver lymphocyte were processed with the Optiprep Nycoprep Lymphoprep Kit (Axis-Shield, Oslo, Norway), and stained with the indicated antibodies for assessment by flow cytometry. For assessment of the cell surface molecules, the anti-CD3 FITC and anti-CD4 APC antibodies (eBioscience, San Diego, CA, USA) were used. The Fixation/Permeabilization Kit (eBioscience) was used to immune-stain the intracellular molecules with anti-IL-22 PE, anti-IL-17 PE and anti-FOXP3 PE (eBioscience). For IL-22 andIL-17 staining, the cells were pretreated using the following procedure: cells were first cultured in DMEM and then stimulated for 6 h with 50ng/mL PMA (Sigma, St. Louis, MO, USA) and 500ng/mL ionomycin (Sigma) in the presence of 1μL/mL GolgiStop (BD Biosciences, San Diego, CA, USA). FACS analyses were carried out with a FACSAria cell sorter (BD Biosciences) and FlowJo analysis software (Tree Star, Ashland, OR, USA).
5
Neutrophil IL-22 Production in Leishmaniasis
6
Comprehensive Flow Cytometry Analysis of Immune Cells
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For flow cytometry of immune cells, lamina propria immune cells were isolated as described.83 (link) For analysis of myeloid immune cells, the cells were washed in PBS, incubated with FcR blocking antibody (Miltenyi Biotec, Bergisch Gladbach, Germany) for 10 minutes, and stained with anti-CD45–Pacific Blue, anti-CD3–BV650, anti-NK1.1–BV650, anti-B220–BV650, anti-CD11b–BV605, anti-CD11c–PECy7, anti-Ly6C–PerCPCy5.5, anti-F4/80–APC, anti-CD64–PE, and anti–MHC-II–AF700 (all from BioLegend, San Diego, CA) for 15–30 minutes. ZOMBI-NIR live dead stain (BioLegend) was used for discrimination between live and dead cells. For cytokine staining, the cells were incubated with ionomycin and PMA in the presence of Brefeldin A for 3.5 hours before surface staining with anti-CD25–AlexaFluor700, anti-CD3–PerCPCy5.5, anti-CD4–BV510, and anti-CD8–BV570 for 15 minutes. Cells then were fixed with the FoxP3 staining kit (eBioscience) according to the manufacturer’s instructions, stained with anti-FoxP3–Pacific Blue, anti–IFN-γ–PECy7, anti-IL17–APC, anti-TNFα–BV650, and anti-IL22–PE for 30 minutes, washed in PermWash buffer (eBioscience), samples were acquired on an LSRII cytometer (BD, Franklin Lakes, NJ), and analyzed using FlowJo (Tree Star, Inc, Ashland, OR). Gating strategy for T and myeloid cells was performed as described previously.32
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