Pe rat anti mouse il 17a

Manufactured by BD

Sourced in China, Switzerland, United States

PE Rat Anti-Mouse IL-17A is a lab equipment product that recognizes the mouse Interleukin-17A (IL-17A) protein. It is a phycoerythrin (PE)-conjugated monoclonal antibody.

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Lab products found in correlation

Facs lsr 2, by BD (1 mentions) Ifn γ fitc, by BD (1 mentions) Il 4 apc, by BD (1 mentions) Brefeldin a, by BD (1 mentions) Rat anti mouse cd8β eflour 450, by Thermo Fisher Scientific (1 mentions) Hamster anti mouse cd3ε apc cy7, by BD (1 mentions) Rat anti mouse cd4 percp cy5, by BD (1 mentions) Apc rat anti mouse cd25, by BD (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Facsverse, by BD (1 mentions) Ionomycin, by Merck Group (1 mentions) Golgistop, by Merck Group (1 mentions) Fixation permeabilization diluent, by Thermo Fisher Scientific (1 mentions) Rat anti mouse cd4 fitc, by Thermo Fisher Scientific (1 mentions) Fix perm kit, by MultiSciences Biotech (1 mentions) Goat anti mouse ionized calcium binding adaptor molecule 1 iba1 antibody, by Abcam (1 mentions) Rabbit anti mouse ccr2 antibody, by Abcam (1 mentions) Rat mog35 55 peptides, by Biosynth (1 mentions) Isotype control, by BD (1 mentions) Cytofix cytoperm soln kit, by BD (1 mentions)

Spelling variants of this product

IL-17A (54 citations) Anti-IL-17A-PE (25 citations) IL-17A-PE (17 citations) Anti-IL-17A (14 citations) PE anti-mouse IL-17A (9 citations) Mouse anti (2 citations) APC- or PE-conjugated anti-mouse IL-17A rat monoclonal antibody (TC11-18H10; (2 citations)

9 protocols using pe rat anti mouse il 17a

Th17/Treg Phenotyping Assay

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250 μL of cytosol was divided into
five 1.5 mL microcentrifuge tubes of 50 μL each. One tube was
without any reagent, and the other four tubes contained 5 μL
of PerCP-Cy 5.5 rat antimouse CD4 reagent (BD Biosciences, Cat no.
561115), 5 μL of PE rat anti mouse IL-17A (BD Biosciences, Cat
no. 561115), and 5 μL of PE rat anti mouse IL-17A (BD Biosciences,
Cat no. 561115), mouse IL-17A (BD Biosciences, Cat no. 561020), Alexa
Fluor 647 rat antimouse Foxp3, and Th17/Treg phenotyping cocktail
(BD Biosciences, Cat no. 560767). The reaction was carried out for
30 min at room temperature, protected from light. After the reaction
was completed, 1 mL of stain buffer was added, and the solution was
centrifuged at 500g for 5 min to remove the stain,
and the washing procedure was repeated once more. Finally, the cell
precipitate was dissolved in 500 μL of stain buffer, and it
was ready for use.

Koh Y.C., Chang Y.C., Lin W.S., Leung S.Y., Chen W.J., Wu S.H., Wei Y.S., Gung C.L., Chou Y.C, & Pan M.H. (2024). Efficacy and Mechanism of the Action of Live and Heat-Killed Bacillus coagulans BC198 as Potential Probiotic in Ameliorating Dextran Sulfate Sodium-Induced Colitis in Mice. ACS Omega, 9(9), 10253-10266.

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Profiling Lung T Cell Activation and Cytokines

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Activation and cytokine expression profiles of infiltrating T cells were investigated by fluorescence-activated cell-sorting (FACS) analysis. 1 × 10⁶ lung T cells were stimulated with RSV-specific peptide (M2_82–90, 1 μM) (Anaspec, Fremont, CA) or PMA/Ionomycin (50 ng/mL and 1 μg/mL, respectively) for 6 h to assess CD8+ T-cell activation and CD4+ T-cell polarization, respectively. Fresh RPMI media without PMA/Ionomycin or M2 peptide was applied to cells as a negative control. Following the stimulation period, brefeldin A (0.5 ng/mL, BD Biosciences) was added to inhibit protein transport for 2 h before extracellular staining with rat anti-mouse CD8β eFlour 450 (clone: eBioH35-17.2) (eBioscience) hamster anti-mouse CD3ε APC-Cy7 (clone: 145-2C11), and rat anti-mouse CD4 PerCP-Cy5.5 (clone: RM4-5) (BD Biosciences). All cells were treated with cytofixation and permeabilization buffer (BD Biosciences) for 20 min prior to staining with rat anti-mouse IL-17A PE (clone: TC11-18H10.1), IFNγ FITC (clone: XMG1.2) and IL-4 APC (clone: 11B11) (BD Biosciences) for 45 min. Cells were washed two times with PBS containing 2% FBS and 0.1% NaN₃ and analyzed on the FACS LSRII (BD Biosciences). At least 10,000 CD8+ events and 30,000 CD4+ events were acquired to ensure sufficient detection of intracellular IFNγ, IL-4, and IL-17A.

Warren K.J., Simet S.M., Pavlik J.A., DeVasure J.M., Sisson J.H., Poole J.A, & Wyatt T.A. (2016). RSV-specific anti-viral immunity is disrupted by chronic ethanol consumption. Alcohol (Fayetteville, N.Y.), 55, 35-42.

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Multiparametric Flow Cytometric Analysis of Immune Cell Subsets

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The purified spleen and liver cells were counted and their viability assessed using the trypan blue exclusion method.²⁹ Then, the absolute cell counts (2 × 10⁶) and suspension were in the complete RPMI 1640 medium. All cells were detected by BD FACSVerse and analysed by FlowJo software (Treestar, Inc, San Carlos, USA).
For Th1/Th2/Th17 analysis, the cells were stimulated for 4 h with 25 ng/mL PMA (Sigma‐Aldrich), 1 mg/mL Ionomycin (Sigma‐Aldrich) and 0.66 μL/mL Golgistop (Sigma‐Aldrich) and cultured at 37°C in 5% CO₂. The cells were harvested and stained with rat anti‐mouse CD3‐PerCP‐Cy™5.5 (BD Pharmingen, San Diego, USA) and rat anti‐mouse CD4‐FITC (eBioscience, San Diego, USA). After being fixed and permeabilized with FIX&PERM Kit (MultiSciences, Hangzhou, China), the cells were intracellularly stained with rat anti‐mouse IFN‐γ‐PE (eBioscience), rat anti‐mouse IL‐4‐PE (BD Pharmingen) and rat anti‐mouse IL‐17A‐PE (BD Pharmingen).
For Tregs analysis, single‐cell suspension was stained with rat anti‐mouse CD4‐FITC (eBioscience) and rat anti‐mouse CD25‐APC (BD Pharmingen). After being washed, fixed and permeabilized with Fixation/Permeabilization Diluent (eBioscience), then the rat anti‐mouse Fc block (BD Pharmingen) was added into cell suspension and incubated for 15 minutes at 4°C. The cells were intracellularly stained with rat anti‐mouse Foxp3‐PE (BD Pharmingen).

Chang H., He K., Li C., Ni Y., Li M., Chen L., Hou M., Zhou Z., Xu Z, & Ji M. (2020). P21 activated kinase‐1 (PAK1) in macrophages is required for promotion of Th17 cell response during helminth infection. Journal of Cellular and Molecular Medicine, 24(24), 14325-14338.

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Experimental Autoimmune Encephalomyelitis Model Reagents

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Rat MOG35–55 peptides were obtained from Biosynth International (Naperville, IL, USA) and purified by high performance liquid chromatography, and the purity of the peptide was >95%. The sequence of MOG35–55 was MEVGWYRSPFSRVVHLYRNGK. Purified Hamster anti-mouse CD3e, Fluorescein Isothiocyanate (FITC) rat anti-mouse CD4, APC rat Anti-mouse CD8a, APC rat anti-mouse IL-4 and PE rat anti-mouse IL-17A antibodies were purchased from BD Pharmingen (Basel, Switzerland); Goat anti-mouse ionized calcium-binding adaptor molecule-1 (IBA1) antibody was obtained from abcam (Cambridge, UK); Rat anti-mouse CD45, Rabbit anti-mouse CCL2 antibody and CCL2 (rat recombinant) was purchased from AbD Serotec (Raleigh, NC, USA)). FITC rat anti-mouse IFN-γ antibody was purchased from eBioscienc (San Diego, CA, USA); rabbit anti-mouse CCR2 antibody was purchased from Abcam (Cambridge, UK). 2-(1-benzyl-indazol-3-yl) methoxy)-2-methyl propionic acid (bindarit) was synthesized by and obtained from Angelini (Angelini Research Center-ACRAF, Italy).

Ji Z., Fan Z., Zhang Y., Yu R., Yang H., Zhou C., Luo J, & Ke Z.J. (2014). Thiamine Deficiency Promotes T Cell Infiltration in Experimental Autoimmune Encephalomyelitis: the Involvement of CCL2. Journal of immunology (Baltimore, Md. : 1950), 193(5), 2157-2167.

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Th17 Cell Profiling in Murine Tissues

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The lungs and spleens of mice were carefully removed, and a single cell suspension was generated using a 200-mesh sieve. Then the lymphocytes were separated with a mouse lymphocyte separation solution (TBD science, Tianjin, China). Cells from lungs, spleens, and blood were all incubated at 37°C in RM1640 (Thermo Fisher Scientific, Waltham, MA, USA) containing Phorbol 12-myristate 13-acetate/ionomycin mixture and Brefeldin A/monensin mixture (Multisciences, Hangzhou, China). The stimulated cells were washed and incubated with fluorescein isothiocyanate Rat Anti-Mouse CD4 (BD Biosciences). For intracellular staining, Cytofix/Cytoperm Soln Kit (BD Biosciences) was used and the cells were stained with PE Rat Anti-Mouse IL-17A (BD Biosciences), after washing with Wash/Perm solution. Isotype controls (BD Biosciences) were used to determine gates for positive antibody responses. Th17 proportion was analyzed using a flow cytometer (BD Biosciences).

Xu L., Xu R.A., Zhang D., Su S., Xu H., Wu Q, & Li Y. (2018). The changes of expressive levels of IL-17A, STAT3, and RORγt in different invasive pulmonary aspergillosis mice. Infection and Drug Resistance, 11, 1321-1328.

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Detailed Flow Cytometry Protocol

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FACS flow cytometry was products of BD company (Becton, Dickinson and Company, East Rutherford, New Jersey, United States); CD4+CD25+ regulatory T cell mice staining kit mainly contains 3 types of antibody, namely, FTTC-anti-mouse-CD4+, APC-anti-mouse-CD25, and PE-Cy5-anti-mouse-Foxp3. The Th17 cell staining kit (PE Rat Anti-Mouse IL-17A, APC IFN-γ Anti-Mouse, FTTC-anti-mouse-CD3e, and PerCP Rat Anti-Mouse CD4) was obtained from BD Company. The TGF-β1, IL-10, and IL-17 Elisa reagent kit was obtained from Shanghai Ao Biological Company (Shanghai, China); the antibodies were obtained from R&D systems (Minneapolis, USA). The Trizol reagent was bought from Invitrogen Company (California, USA). Foxp3, RORγt, and the reference GAPDH primers were synthesized by Shanghai ShineGene Biotechnology Companies (Shanghai, China); cDNA synthesis kit and PCR amplification kit were provided by TaKaRa Co. Biological Engineering (Dalian, China); protein extraction kit was provided by the Bestbio Biotechnology Companies (Shanghai, China); Mouse Anti-Human Foxp3 (2A11GG) polyclonal antibody and Rabbit Anti-Human RORγt (H-190) polyclonal antibody were purchased from Santa-Cruz Company (Santa Cruz Biotechnology, Inc.). Reference antibody Histone H3.1 (Ab-10) was obtained from SignalWay Antibody (Maryland, USA).

Wang L., Du H., Liu Y., Wang L., Ma X, & Zhang W. (2015). Chinese Medicine Bu Xu Hua Yu Recipe for the Regulation of Treg/Th17 Ratio Imbalance in Autoimmune Hepatitis. Evidence-based Complementary and Alternative Medicine : eCAM, 2015, 461294.

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Murine T Cell Immunophenotyping

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Antibodies for FACS included PBB515 Rat Anti-Mouse CD25 (BD, cat. no.564424), PE-Cy-5 Rat Anti-Mouse CD4 (BD, cat. no.553050), PE-Cy-7 ANTI-M/R Foxp3 FJK-16S (BD, cat. no.25-5773-82), and PE Rat Anti-Mouse IL-17A (BD, cat. no.559502).

Liu M., Mao J, & Zhang S. (2022). Effect of Intervention of Probiotics in Advance on Treg/Th17 in Premature Mice. BioMed Research International, 2022, 6131069.

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Flow Cytometric Analysis of Lymphocyte Subsets

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The ratios of T lymphocyte subsets in the spleen of treated mice were detected with flow cytometry. Spleens were grounded with mortar and pestle in lymphocyte separation medium (Catalog #: DKW33-R0100, Dakewe Biotech Co., Ltd., Shenzhen, China) and then incubated with FITC Rat Anti-Mouse CD4 (Clone RM4-5, Catalog #: 553046, BD Biosciences, Franklin Lakes, NJ, USA)/APC Rat Anti-Mouse CD25 (Clone PC61, Catalog #: 561048, BD Biosciences, Franklin Lakes, NJ, USA)/PE Anti-Mo/Rt Foxp3 (Clone FJK-16a, Catalog #: 2344844, Elabscience Biotechnology, Wuhan, China)/APC Rat Anti-Mouse CD3 (Catalog #: 554832, BD Biosciences, Franklin Lakes, NJ, USA)/PE Rat Anti-Mouse IL-17A (Catalog #: 553046, BD Biosciences, Franklin Lakes, NJ, USA) and analyzed with a Cytoflex Platform (Cytoflex S, Beckman Coulter Life Sciences, Indianapolis, IN, USA). The percentage of CD4+IL-17A+ Th17 cell and CD4+CD25+Foxp3+ Treg cells in total T cells were analyzed by using CellQuest™ 2.1 software (BD, San Diego, CA, USA).

Wang M., Su T., Sun H., Cheng H., Jiang C., Guo P., Zhu Z., Fang R., He F., Ge M., Guan Q., Wei W, & Wang Q. (2022). Regulating Th17/Treg Balance Contributes to the Therapeutic Effect of Ziyuglycoside I on Collagen-Induced Arthritis. International Journal of Molecular Sciences, 23(24), 16105.

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Comprehensive Immune Cell Profiling by Flow Cytometry

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Flow cytometry was performed on a CytoFLEX (Beckman), and data were analyzed by FlowJo software. The live cells were observed by the Fixable Viability Stain 780 (Cat#565388, BD Pharmingen). For cell-surface staining, cells were stained with BV510 Rat Anti-Mouse CD45(Cat#563891, BD Pharmingen), FITC Hamster Anti-Mouse CD3e (Cat#553061, BD Pharmingen), PE-Cy7 Rat Anti-Mouse CD4 (Cat#552775, BD Pharmingen), BV421 Rat Anti-Mouse CD25 (Cat#562606, BD Pharmingen), FITC Rat Anti-CD11b (Cat#557396, BD Pharmingen), BV421 Rat Anti-Mouse F4/80 (Cat#565411, BD Pharmingen), PE Rat Anti-Mouse CD86(Cat#561963 BD Pharmingen), Alexa Fluor 647 Rat Anti-Mouse CD206 (Cat#565250 BD Pharmingen). For intracellular staining, cells were stained with PE Rat Anti-Mouse IL-17A (Cat#561020 BD Pharmingen), Alexa Fluor 647 Rat anti-Mouse Foxp3(Cat#560402 BD Pharmingen). According to the reagent vendor’s instructions, Purified Rat Anti-Mouse CD16/CD32 is used to exclude background fluorescent signal interference, Cells fixation and permeabilization Fixation/Permeablization Kit (Cat#554714 BD Pharmingen) and Transcription Factor Buffer Set (Cat#562574 BD Pharmingen) were used for cell fixation and permeabilization.

Li C., Liu Y., Deng M., Li J., Li S., Li X., Zuo Y., Shen C, & Wang Y. (2024). Comparison of the therapeutic effects of mesenchymal stem cells derived from human dental pulp (DP), adipose tissue (AD), placental amniotic membrane (PM), and umbilical cord (UC) on postmenopausal osteoporosis. Frontiers in Pharmacology, 15, 1349199.

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