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Human mouse tgf β1 elisa ready set go kit

Manufactured by Thermo Fisher Scientific
Sourced in United States

The Human/Mouse TGF-β1 ELISA Ready-Set-Go kit is a laboratory assay used to measure the concentration of TGF-β1 in human and mouse biological samples. The kit provides the necessary components for performing a sandwich enzyme-linked immunosorbent assay (ELISA) to detect and quantify TGF-β1 levels.

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5 protocols using human mouse tgf β1 elisa ready set go kit

1

Cytokine Profiling in Pancreatic Tissue

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Pieces of pancreas were removed, weighed, and placed in a tube containing 700 μl Complete Protease Inhibitor Cocktail (Roche Diagnostics). Pancreatic tissue was homogenized using a Polytron homogenizer (Kinematica, Luzern, Switzerland) and IL-2, IL-6, IFNγ, TNFα, IL-17, IL-4, and IL-10 levels were detected by the cytometric bead array (CBA) (Th1/Th2/Th17 kit; BD) method, according to the manufacturer’s instructions. The concentration of TGF-β in pancreatic tissue was determined using Human/Mouse TGF-β1 ELISA Ready-Set-Go kit (eBioscience, San Diego, CA, USA). Serum cytokine levels were also determined by the CBA method.
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2

Quantifying Transforming Growth Factor-β1

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Concentrations of TGF-β1 were determined using a Human/Mouse TGF-β1 ELISA Ready-Set-Go kit (eBioscience, San Diego, CA) according to the manufacturer’s protocol (see Online Supplementary Materials for details). The samples were tested in triplicate. Human TGF-β1 standards were run in duplicate two-fold dilutions ranging from 1,000 to 15.6 pg/ml. The plate was read at 450 and 560 nm. The TGF-β1 OD values were obtained by subtraction of the 560 nm background values from the 450 nm values. The concentration of TGF-β1 was calculated based on the TGF-β1 standard curve. Data were normalized by calculating the concentration of TGF-β1 in picograms per milligram of total protein.
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3

Quantifying Serum TGF-β in NOD Mice

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TGF-β levels were quantified in serum at 4 and 6 weeks (5 mice/time-point) in MK626-treated and control NOD mice, using a specific Human/Mouse TGF-β1 ELISA Ready-SET-Go kit (e-Bioscience, USA). The experiment was replicated twice.
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4

Measuring TGF-β Activation in Pulmonary Fibrosis

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Activation of TGF-β in cell culture experiments or in BALF obtained from WT and Del-1−/− mice with PF induced by inactive TGF-β-expressing adenovirus was measured using the human/mouse TGF-β1 ELISA Ready-SET Go kit (eBioscience) according to the manufacturer's protocol. Briefly, the plate was coated overnight at 4°C with capture antibody, washed five times with PBST (0.05% Tween 20 in PBS), blocked with 1× assay buffer, incubated at RT for 1 h, and washed five times with 0.05% PBST. The plate was then loaded with samples and standards, incubated at RT for 2 h or at 4°C overnight, washed five times with 0.05% PBST, and incubated at RT for 1 h with biotin-conjugated anti-active TGF-β antibody. After washing five times with 0.05% PBST, the plate was incubated at RT for 30 min with avidin-conjugated HRP. The plate was washed five times with 0.05% PBST, developed with TMB, and read at 630 nm on a microplate reader (Synergy HT). The concentrations of LAP were measured as described above but using a mouse LAP (TGF beta 1) Ready-SET Go kit (eBioscience).
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5

Multiplex Analysis of Pancreatic Cytokines

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Pieces of pancreas were removed, weighed and placed into a tube containing Complete Protease Inhibitor Cocktail (Roche Diagnostics, Abbott Park, IL, USA). Pancreatic tissue was homogenized using a Polytron homogenizer (Kinematica, Luzern, Switzerland) and interleukin (IL)-2, IL-6, interferon gamma, IL-17, IL-4 and IL-10 levels were detected in the supernatant by the cytometric bead array method (Th1/Th2/Th17 kit; BD), according to the manufacturer’s instructions. The concentration of transforming growth factor beta (TGF-β) in pancreatic tissue was determined using the Human/Mouse TGF-β1 ELISA Ready-Set-Go kit (eBioscience, San Diego, CA, USA). Serum cytokine levels were also determined by the cytometric bead array method.
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