Odyssey imager

Manufactured by Thermo Fisher Scientific

The Odyssey Imager is a high-performance fluorescence imaging system designed for a wide range of applications in life science research. It utilizes near-infrared fluorescence detection to enable quantitative and sensitive imaging of proteins, nucleic acids, and small molecules.

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Odyssey Fc Imager (3 citations)

3 protocols using odyssey imager

Western Blotting for Protein Expression

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For western blotting, 20–40 μg of total protein from each sample was subjected to SDS-PAGE under reducing conditions. Proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore). The membranes were blocked for 1 h at room temperature and then incubated overnight at 4 °C with primary antibodies. Primary antibodies used were rabbit anti-COX-2 (Santa Cruz), rabbit anti-iNOS (Santa Cruz), rabbit anti-acetyl-p65 (Cell Signalling), rabbit anti-Total-p65 (Cell Signalling), rabbit anti-MAP2(Cell Signalling), rabbit anti-Nrf2 (Santa Cruz), rabbit anti-HO1 (Santa Cruz), rabbit anti-NQO1 (Santa Cruz), rabbit anti-SIRT1 (Santa Cruz) and rabbit anti-actin (Sigma). Primary antibodies were diluted in Tris-buffered saline (TBS), containing 0.1% Tween 20 (TBS-T) and 1 or 5% BSA. Membranes were incubated with the primary antibody overnight at 4 °C. After extensive washing (three times for 15 min each in TBS-T), proteins were detected by incubation with Alexa Fluor 680 goat anti-rabbit secondary antibody (1:10,000; Life Technologies) at room temperature for 1 h. Detection was done using a LICOR Odyssey Imager. All western blot experiments were carried out at least three times.

Velagapudi R., El-Bakoush A, & Olajide O.A. (2018). Activation of Nrf2 Pathway Contributes to Neuroprotection by the Dietary Flavonoid Tiliroside. Molecular Neurobiology, 55(10), 8103-8123.

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Western Blotting for Protein Expression

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For Western blotting, 20-40 μg of total protein from each sample was subjected to SDS-PAGE under reducing conditions. Proteins were then transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore). The membranes were blocked for 1 h at room temperature and then incubated overnight at 4°C with primary antibodies. Primary antibodies used were rabbit anti-COX-2 (Santa Cruz), rabbit anti-iNOS (Santa Cruz), rabbit anti-phospho-IκBα (Santa Cruz), rabbit antiphospho-p65 (Cell Signalling), rabbit anti-p65 (Cell Signalling) rabbit anti-acetyl-p65 (Cell Signalling), rabbit anti-Nrf2 (Santa Cruz), rabbit anti-HO1 (Santa Cruz), rabbit anti-NQO1 (Santa Cruz), and rabbit anti-actin (Sigma). Primary antibodies were diluted in Tris-buffered saline (TBS), containing 0.1% Tween 20 (TBS-T) and 1 or 5% BSA. Membranes were incubated with the primary antibody overnight at 4°C. After extensive washing (three times for 15 min each in TBS-T), proteins were detected by incubation with Alexa Fluor 680 goat anti-rabbit secondary antibody (1:10000; Life Technologies) at room temperature for 1 h. Detection was done using a LICOR Odyssey Imager. All Western blot experiments were carried out at least three times.

Velagapudi R., Kumar A., Bhatia H.S., El-Bakoush A., Lepiarz I., Fiebich B.L, & Olajide O.A. (2017). Inhibition of neuroinflammation by thymoquinone requires activation of Nrf2/ARE signalling. International immunopharmacology, 48.

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Western Blot Analysis of Inflammatory Signaling Proteins

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Equal amounts of protein (20 μg) were separated on a polycryamide electrophoresis gel and transferred onto a polyvinylidine fluoride (PVDF) membrane. Membranes were incubated in blocking buffer for 1 h at room temperature, washed 3 times for 10 min each in Tris-buffered saline containing 0.1% Tween 20 (TBS-T), and incubated with primary antibodies overnight at 4C. Primary antibodies used were rabbit anti-COX-2 (Santa Cruz;
1:500), rabbit anti-iNOS (Santa Cruz, 1:500), rabbit anti-IBα (Santa Cruz; 1:250), rabbit anti-phospho-IκBα (Santa Cruz, 1:250), rabbit anti-IKKα (Santa Cruz; 1:250), rabbit antiphospho-IKKα (Santa Cruz, 1:250), rabbit anti-phospho-p65 (Santa Cruz, 1:500), rabbit anti-p65 (Santa Cruz 1:500), rabbit anti-microtubule-associated protein-2 (MAP2) (Santa Cruz, 1:500) , rabbit anti-ERβ (Santa Cruz 1:250) and rabbit anti-actin (Sigma Aldrich, 1:500). Primary antibodies were diluted in TBS-T and 1% bovine serum albumin (BSA).
After extensive washing (three times for 15 min each in TBS-T), proteins were detected by incubation with Alexa Fluor 680 goat anti-rabbit secondary antibody (1:10000; Life Technologies) at room temperature for 1 h. Detection was done using a LICOR Odyssey Imager. All Western blot experiments were carried out at least three times.

El-Bakoush A, & Olajide O.A. (2018). Formononetin inhibits neuroinflammation and increases estrogen receptor beta (ERβ) protein expression in BV2 microglia. International immunopharmacology, 61.

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