Odyssey clx near infrared imager

Manufactured by LI COR

The Odyssey CLx near-infrared imager is a versatile laboratory instrument used for imaging and quantifying fluorescent and chemiluminescent signals. It features high sensitivity, dynamic range, and resolution to provide accurate and reliable data for a wide range of applications.

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Lab products found in correlation

Ab110411, by Abcam (3 mentions) Trans blot turbo transfer system, by Bio-Rad (3 mentions) Intercept tbs blocking buffer, by LI COR (1 mentions) C raf, by Cell Signaling Technology (1 mentions) B raf, by Cell Signaling Technology (1 mentions) Mek1 2, by Cell Signaling Technology (1 mentions) Erk1 2, by Cell Signaling Technology (1 mentions) Ab73400, by Abcam (1 mentions) Irdye 800cw goat anti rabbit, by LI COR (1 mentions) Image studio software, by LI COR (1 mentions) Irdye 800cw conjugated goat anti human secondary antibody, by LI COR (1 mentions) Blocking buffer, by LI COR (1 mentions) Novex sharp pre stained protein standard, by Thermo Fisher Scientific (1 mentions) Ab186734, by Abcam (1 mentions) A2172, by Merck Group (1 mentions) Tgx gel, by Bio-Rad (1 mentions) Sc 98900, by Santa Cruz Biotechnology (1 mentions) Sc 33796, by Santa Cruz Biotechnology (1 mentions) Rc dc kit, by Bio-Rad (1 mentions) Sc 8828, by Santa Cruz Biotechnology (1 mentions)

6 protocols using odyssey clx near infrared imager

Immunoblotting of MAPK Pathway Proteins

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Immunoblotting was performed as previously described (Brian et al., 2022 (link); Brian et al., 2020 (link)). Briefly, 0.025 million cell equivalents of whole cell lysate were run through 7% Tris-acetate polyacrylamide gels and transferred to polyvinylidene difluoride membranes. Membranes were blocked in Intercept (TBS) Blocking Buffer (LI-COR Biosciences) for 1 hr, and then incubated overnight with primary antibody: ARAF (Cell Signaling), BRAF (Cell Signaling), CRAF (Cell Signaling), MEK1/2 (Cell Signaling), and ERK1/2 (Cell Signaling). Membranes were washed and incubated for 1 hr at room temperature with corresponding species-reactive secondary antibody and then imaged using an Odyssey CLx near-infrared imager (LI-COR Biosciences).

Rasmussen D.M., Semonis M.M., Greene J.T., Muretta J.M., Thompson A.R., Toledo Ramos S., Thomas D.D., Pomerantz W.C., Freedman T.S, & Levinson N.M. (2024). Allosteric coupling asymmetry mediates paradoxical activation of BRAF by type II inhibitors. eLife, 13, RP95481.

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Western Blot Analysis of Osteogenic Markers

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The cells were cultured as described in the ALP assay. The harvested cell lysates after 3 days were measured using a BCA protein assay. Equal amounts of proteins (15–30 μg) were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membranes, before immunoblotting with rabbit anti-Runx2 (D1L7F, CST, Danvers, MA, USA), rabbit anti-OPG (ab73400, Abcam, Cambridge, MA, USA), and mouse anti-RANKL (12A668, Abcam) primary antibodies. The rabbit anti-α/β-tubulin (2148, Abcam) was used as a loading control. Goat anti-rabbit (IRDye 800CW) and goat anti-mouse (IRDye 680CW) fluorochrome-conjugated secondary antibodies were purchased from Licor Biosciences (Lincoln, NB, USA). The protein bands were imaged on an Odyssey CLx near-infrared imager (Licor Biosciences) and quantified by Image Studio software (Licor Biosciences). Each band was normalized to its corresponding loading control band. The results were expressed as the percentage change compared to the untreated control.

Bu T., Ren Y., Yu S., Zheng J., Liu L., Sun P., Wu J, & Yang K. (2022). A Low-Phenylalanine-Containing Whey Protein Hydrolysate Stimulates Osteogenic Activity through the Activation of p38/Runx2 Signaling in Osteoblast Cells. Nutrients, 14(15), 3135.

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SDS-PAGE and Western Blot Analysis of B1 Clade Virus

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B1 clade virus preparation was mixed with denaturing and reducing loading buffer, boiled at 100°C for 5 min, and then run on an SDS-PAGE gel along with Novex sharp pre-stained protein standard (ThermoFisher Scientific). Protein was transferred to a membrane, blocked in blocking buffer (Li-Cor), and then cut into strips so that individual lanes could be stained with purified mAb in blocking buffer. An IRDye 800CW-conjugated goat anti-human secondary antibody (Li-Cor) was used to detect mAb binding. Strips were reassembled to visualize molecular weight and imaged on an Odyssey CLx near infrared imager (Li-Cor).

Vogt M.R., Fu J., Kose N., Williamson L.E., Bombardi R., Setliff I., Georgiev I.S., Klose T., Rossmann M.G., Bochkov Y.A., Gern J.E., Kuhn R.J, & Crowe JE J.r. (2020). Human Antibodies Neutralize Enterovirus D68 and Protect Against Infection and Paralytic Disease. Science immunology, 5(49), eaba4902.

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Mitochondrial Protein Analysis in Human Muscle

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Western blot analyses were performed in RIPA-lysates of human muscle tissue. Protein concentration was determined using the Bio-Rad RC/DC kit (Bio-Rad Laboratories, Veenendaal, The Netherlands). Equal amounts of protein were loaded on 12% TGX gels (Bio-Rad Laboratories) or 4–12% Bolt gradient gels (Novex, Thermo Fisher Scientific, Bleiswijk, The Netherlands). Proteins were transferred to nitrocellulose with the Trans-Blot Turbo transfer system (Bio-Rad Laboratories). Primary antibodies: a cocktail of mouse monoclonal antibodies directed against human OXPHOS (dilution 1:10,000; ab110411, Abcam, Cambridge, UK), two mitochondrial markers directed against TOMM20 (dilution 1:10,000; ab186734; Abcam), porin/VDAC (dilution 1:10,000; sc-8828, Santa Cruz Biotechnology, Dallas, Texas), SR-actin (dilution 1:5,000; A-2172; Sigma Aldrich, Zwijndrecht, The Netherlands), PGC-1 (dilution 1:10,000, 516,557, Calbiochem), FIS-1 (dilution 1:1000, sc-98900, Santa Cruz Biotechnology, Dallas, Texas), PINK-1 (dilution 1:2000, sc-33796, Santa Cruz Biotechnology, Dallas, Texas) and OPA-1 (dilution 1:2500, 612,606, Becton Dickinson). The specific proteins were detected using secondary antibodies conjugated with IRDye680 or IRDye800, and were quantified with the CLx Odyssey Near Infrared Imager (Li-COR, Westburg, Leusden, The Netherlands).

van Moorsel D., Hansen J., Havekes B., Scheer F.A., Jörgensen J.A., Hoeks J., Schrauwen-Hinderling V.B., Duez H., Lefebvre P., Schaper N.C., Hesselink M.K., Staels B, & Schrauwen P. (2016). Demonstration of a day-night rhythm in human skeletal muscle oxidative capacity. Molecular Metabolism, 5(8), 635-645.

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Mitochondrial OXPHOS Protein Quantification

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Mitochondrial content was assessed by mitochondrial OXPHOS protein expression using western blot analyses in Bioplex-lysates of human muscle tissue as previously described^{62 (link)}. Equal amounts of protein were loaded 4–12% Bolt gradient gels (Novex, Thermo Fisher Scientific, Bleiswijk, The Netherlands). Proteins were transferred to nitrocellulose with the Trans-Blot Turbo transfer system (Bio-Rad Laboratories). Primary antibodies contained a cocktail of mouse monoclonal antibodies directed against human OXPHOS (dilution 1:10,000; ab110411, Abcam, Cambridge, UK). The hOxPHOS proteins were detected using secondary antibodies conjugated with IRDye680 or IRDye800 and were quantified with the CLx Odyssey Near-Infrared Imager (Li-COR, Westburg, Leusden, The Netherlands).

Grevendonk L., Connell N.J., McCrum C., Fealy C.E., Bilet L., Bruls Y.M., Mevenkamp J., Schrauwen-Hinderling V.B., Jörgensen J.A., Moonen-Kornips E., Schaart G., Havekes B., de Vogel-van den Bosch J., Bragt M.C., Meijer K., Schrauwen P, & Hoeks J. (2021). Impact of aging and exercise on skeletal muscle mitochondrial capacity, energy metabolism, and physical function. Nature Communications, 12, 4773.

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Western Blot Analysis of OXPHOS Proteins

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Western blot analyses were performed in Bioplex‐lysates of human muscle tissue as previously described (Wefers et al., 2020 (link)). Equal amounts of proteins were loaded on gradient Bolt 4%–12% gels (Novex, Thermo Fisher Scientific). Proteins were transferred to nitrocellulose with the Trans‐Blot Turbo transfer system (Bio‐Rad Laboratories). The following antibodies and dilutions were used in this study: a cocktail of mouse monoclonal antibodies directed against human OXPHOS (dilution 1:5.000; ab110411, Abcam), as well as antibodies directed against TOMM20 (dilution 1:10.000; aba186734; Abcam), porin/VDAC (dilution 1:1.000; sc‐390,996; 1:5000, Santa Cruz biotechnology). The specific proteins were detected using secondary antibodies conjugated with IRDye680 or IRDye800 and were quantified with the CLx Odyssey Near Infrared Imager (Li‐COR, Westburg).

Mancilla R., Pava‐Mejia D., van Polanen N., de Wit V., Bergman M., Grevendonk L., Jorgensen J., Kornips E., Hoeks J., Hesselink M.K, & Schrauwen‐Hinderling V.B. (2023). Invasive and noninvasive markers of human skeletal muscle mitochondrial function. Physiological Reports, 11(12), e15734.

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