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Odyssey p140 clx infrared imaging system

Manufactured by LI COR
Sourced in United States

The Odyssey P140-CLx Infrared Imaging System is a multi-channel fluorescence imaging system designed for high-performance Western blot and in-gel fluorescence detection. The system provides a wide dynamic range and supports multiple applications, including Western blotting, plate-based assays, and in-gel fluorescence analysis.

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3 protocols using odyssey p140 clx infrared imaging system

1

Quantitative Western Blot Analysis of Tau, Amyloid, and NADPH Oxidase Proteins in Mouse Brain

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Mouse brain tissue was homogenized in lysis buffer containing 50 mM Tris-HCL (pH 7.4), 100 mM NaF, 2 mM EDTA, 10 mM β-mercaptoethanol, 2 mM NaVanadate, 8.5% sucrose, 5 μg/ml aprotinin, 100 μg/ml leupeptin, and 5 μg/ml pepstatin. Protein concentrations were determined by using BCA Kits according to the manufacturer’s protocol. Quantitative homogenates were added 5 × sodium dodecyl sulfate (SDS) to heat for 10 min at 99 °C and resolved in 10% SDS-PAGE for separation. After separated, samples were transferred onto nitrocellulose membranes (GE Healthcare, Wauwatosa, WI), and the membranes were blocked with 5% non-fat milk for 1 h at room temperature. Then, the membranes were incubated with primary antibodies including Tau5 (1:1000), Tau1 (1:1000), the anti-phospho-tau antibodies pS199 (1:1000), pT205 (1:1000), pS396 (1:1000), pS404 (1:1000), anti-β-Amyloid (6E10, 1:2000), anti-APP (1:20000), anti-p47phox (1:250), anti-p67phox (1:1000), anti-NOX2/gp91phox (1:1000), anti-GFAP (1:1000), anti-GAPDH (1:20000), and anti-β-Actin (1:20000), followed by the respective IRDye® 800CW or IMDye® 800CW secondary antibodies. The membranes were scanned using an Odyssey P140-CLx Infrared Imaging System (LI-COR, Inc.). Densitometric quantification of protein bands was analyzed using the ImageJ software (National Institutes of Health, Bethesda, MD).
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2

Western Blot Analysis of Mouse Brain Proteins

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Mouse brain tissue was homogenized in lysis buffer containing 50 mM Tris-HCl (pH 7.4), 100 mM NaF, 2 mM EDTA, 10 mM β-mercaptoethanol, 2 mM NaVanadate, 8.5% sucrose, 5 μg/mL aprotinin, 100 μg/mL leupeptin, and 5 μg/mL pepstatin. Protein concentrations were determined using a BCA Kit according to the manufacturer’s protocol. Then, the tissue homogenates were heated in 5× sodium dodecyl sulfate (SDS)-PAGE loading buffer at 99 °C for 10 min. Tissue homogenates were separated on 10% SDS-PAGE, and separated samples were transferred onto nitrocellulose membranes (Whatman Protran®, Sigma). The membranes were blocked with 5% non-fat milk for 1 h at room temperature and incubated overnight at 4 °C with primary antibodies including anti-GFAP (1:1000) and anti-GAPDH (1:1000), followed with corresponding secondary antibodies IRDye® 800CW or IMDye® 800CW antibodies for 1 h at room temperature. The membranes were scanned using the 800-nm channel of an Odyssey® P140-CLx Infrared Imaging System (LI-COR, Inc.). Densitometric quantification of protein bands was analyzed using the ImageJ software (National Institute of Mental Health, Bethesda, MD).
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3

Western Blotting Analysis of Mouse Brain Proteins

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The mouse brain tissues were homogenized in lysis buffer including 50 mM Tris-HCL (pH 7.4), 100 mM NaF, 2 mM EDTA, 10 mM β-mercaptoethanol, 2 mM NaVanadate, 8.5% sucrose, 5 μg/mL aprotinin, 100 μg/mL leupeptin, and 5 μg/mL pepstatin. The protein concentrations were detected using BCA Kits according to the manufacturer’s protocol. The quantitative homogenates were added to a 5× sodium dodecyl sulfate (SDS)-PAGE loading buffer, heated for 10 min at 99 °C, and separated onto 10% or 12% SDS-PAGE. After separation, the samples were transferred onto nitrocellulose membranes (GE Healthcare, Wauwatosa, WI, USA), and the membranes were blocked with 5% non-fat milk for 1 h at room temperature. Then, the membranes were incubated with primary antibodies including anti-Iba1 (1:1000), anti-chemerin (1:1000), anti-CMKLR1 (1:1000), and anti-GAPDH (1:20,000), followed by IRDye® 800CW or IMDye® 800CW secondary antibodies. The membranes were scanned using an Odyssey P140-CLx Infrared Imaging System (LI-COR, Inc., Lincoln, NE, USA). Densitometric quantification of protein bands was analyzed by the ImageJ software (National Institutes of Health, Bethesda, MD, USA).
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