SIC and LNC were stimulated with PMA and ionomycin as described above and then incubated with 10% normal rat serum for 15 min. The T cells among SIC and LNC were surface-stained with anti-CD3 APC, anti-CD4 FITC, and/or anti-CD25 PE (all from eBioscience, San Diego, CA, USA).22 (link) For intracellular staining, cells were fixed and permeabilized using the Fixation/ Permeabilization Kit (eBioscience, San Diego, CA, USA) followed by staining with anti-rat/mouse IL-17A eFlour 450 (eBioscience, San Diego, CA, USA) and/or anti-rat IFNγ (BioLegend, San Diego, CA, USA).22 (link) Foxp3 was stained using anti-mouse/rat Foxp3 eFlour 450 (eBioscience). Using LSRII flow cytometer (BD Bioscience) (Flow Cytometry Shared Service, UMB), lymphocytes were acquired after setting “gates” for size (forward scatter) and cell complexity (side scatter) and further gated for CD3+ and CD4+ T cells. Thereafter, CD4+IL-17A+ gates and CD25+Foxp3+ gates were used to quantify Th17 and Treg cell frequency, respectively. Th17/Treg ratio was calculated using percentage of cell numbers. Appropriate isotype controls were used to establish each positive signal. Data analysis was performed using FlowJo software (Tree Star).
Anti rat ifnγ
Manufactured by BioLegend
Sourced in United States
Anti-rat IFNγ is a laboratory reagent used to detect and quantify the presence of interferon-gamma (IFNγ), a cytokine secreted by various immune cells, in rat biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd3 apc, by Thermo Fisher Scientific (1 mentions)
Anti cd4 fitc, by Thermo Fisher Scientific (1 mentions)
Anti cd25 pe, by Thermo Fisher Scientific (1 mentions)
Fixation permeabilization kit, by Thermo Fisher Scientific (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Alexa fluor 647, by BioLegend (1 mentions)
Fitc anti rat cd4, by BioLegend (1 mentions)
Pe anti rat cd25, by BD (1 mentions)
Rat il 4 elisa kit, by Thermo Fisher Scientific (1 mentions)
Rabbit anti rat cd68, by Abcam (1 mentions)
Rabbit anti rat cd103, by Abcam (1 mentions)
Pe anti rat il 4, by BioLegend (1 mentions)
Lipase, by Nanjing Jiancheng (1 mentions)
D lactate, by Nanjing Jiancheng (1 mentions)
2 protocols using anti rat ifnγ
1
Th17 and Treg Cell Quantification
2
Multiplex Biomarker Quantification in Rat
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Amylase, lipase, D-lactate, soluble tumor necrosis factor-α receptor (sTNF-αR), and secretory immunoglobulin A (sIgA) enzyme-linked immunosorbent assay (ELISA) kits were purchased from Nanjing Jiancheng Biological Engineering Institute (Nanjing, China). The chromogenic limulus amebocyte lysate kit was purchased from Bioendo Technology Co., Ltd (Xiamen, China). Rat interleukin-1beta (IL-1β) and transforming growth factor-β (TGF-β) ELISA kits were purchased from ExCell Biotech Co., Ltd (Shanghai, China). Rat TNF-α and IL-10 ELISA kits were purchased from Cusabio Biotech Co., Ltd (Wuhan, China). Rat IL-6 and IL-18 ELISA kits were purchased from Elabscience Biotech Co., Ltd (Wuhan, China). The rat IL-8 ELISA kit was purchased from Neobioscience Technology Co., Ltd (Shenzhen, China). The rat IL-4 ELISA kit was purchased from Invitrogen (Carlsbad, CA, USA). The specific antibodies against FITC anti-rat CD4, Alexa fluor®647, anti-rat IFN-γ, PE anti-rat IL-4, and Alexa fluor® 647 anti-rat Foxp3 were purchased from BioLegend (San Diego, CA, USA). Rabbit anti-rat CD68 and rabbit anti-rat CD103 were purchased from Abcam (Cambridge, MA, UK). PE anti-rat CD25 was purchased from BD Biosciences (San Jose, CA, USA). The sources of other reagents were indicated in the specified methods.
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