Odyssey western blot detection system

Manufactured by LI COR

The Odyssey Western Blot Detection System is a fluorescence-based imaging system designed for the detection and quantification of proteins in Western blot analysis. The core function of the system is to capture and analyze near-infrared fluorescent signals from labeled protein targets on a membrane.

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Lab products found in correlation

Complete protease inhibitor cocktail, by Roche (1 mentions) Rabbit anti irisin antibody, by Phoenix Pharmaceuticals (1 mentions) Rabbit anti cytochrome c antibody, by Abcam (1 mentions) Anti rabbit secondary antibody, by LI COR (1 mentions) T stat3, by Abcam (1 mentions) Rorγt, by Abcam (1 mentions) T enos, by Cell Signaling Technology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) P stat3, by Abcam (1 mentions)

3 protocols using odyssey western blot detection system

Quantitative Western Blot Analysis

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Cell lysates were prepared using KALB lysis buffer (150 mM NaCl, 50 mM Tris (base), pH 7.4, 1 mM EDTA, 1% Triton X-100 and 10% glycerol) and complete protease inhibitor cocktail (Roche). Proteins were resolved using SDS–PAGE on a Bis-Tris 4–12% gradient gel (Invitrogen). Protein bands were visualized using the Odyssey western blot detection system (Li-Cor Biosciences) (Supplementary Fig. 1a). Densitometry was performed using ImageJ (National Institutes of Health) where the relative fluorescence intensity of each protein band was calculated relative to that of GAPDH.

Lee N.K., Fok K.W., White A., Wilson N.H., O'Leary C.J., Cox H.L., Michael M., Yap A.S, & Cooper H.M. (2016). Neogenin recruitment of the WAVE regulatory complex maintains adherens junction stability and tension. Nature Communications, 7, 11082.

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Immunoblotting Analysis of Irisin, UCP2, Caspase 9, and Cytochrome c

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Lung tissues were washed twice with ice-cold PBS and lysed in lysis buffer [10mM tris-HCl (pH 8.0), 150 mM NaCl, 1% NP-40, 1mM phenylmethylsulfonyl fluoride, and leupeptin and aprotinin (10 mg/ml each)]. After centrifugation at 12,000g for 15 min, the supernatants were collected and their protein concentrations were measured using a BCA protein assay kit (HyClone Pierce). The tissue homogenates (50 μg of protein) were separated by 10% SDS-PAGE and transferred onto nitrocellulose membranes. The blots were then washed with tris-buffered saline with Tween 20 (TBST), blocked with 5% milk powder in TBST buffer for 1 hour, and incubated with the appropriate primary antibodies at appropriate dilutions. The blotted membranes were probed with the rabbit anti-irisin antibody (1:500; Phoenix Pharmaceuticals), the rabbit anti-UCP2 antibody (1:500; Sigma-Aldrich), rabbit anti-active caspase 9 antibody (1:500; Abcam), and rabbit anti–cytochrome c antibody (1:500, Abcam) at 4°C overnight. Then, the membranes were washed and primary antibodies were detected with fluorescent-labeled goat anti-rabbit IgG (1:15,000; IRDye, LI-COR), and the bands were visualized by Odyssey Western Blot Detection System (LI-COR). The amount of protein transferred onto the membranes was verified by immunoblotting for glyceraldehyde-3-phosphate dehydrogenase.

Chen K., Xu Z., Liu Y., Wang Z., Li Y., Xu X., Chen C., Xia T., Liao Q., Yao Y., Zeng C., He D., Yang Y., Tan T., Yi J., Zhou J., Zhu H., Ma J, & Zeng C. (2017). Irisin protects mitochondria function during pulmonary ischemia/reperfusion injury. Science translational medicine, 9(418), eaao6298.

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Western Blotting of Vascular Proteins

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Western blotting was performed according to standard protocols described previously (Citation15). Briefly, thoracic aortas and CD4 + T cells were homogenized using icecold RIPA lysis buffer. The proteins lysates were separated by SDS-PAGE and transferred onto nitrocellulose membranes. The membranes were blocked using 8% fat-free milk in TBS (Tris-buffered saline) with 0.5% Tween-20 for 2 h at room temperature and then probed with primary antibodies against phosphorylated eNOS at Ser 1177 (1:500; Cell Signaling Technology, Danvers, MA, USA), t-eNOS (1:1000; Cell Signaling Technology), p-STAT3 (1:400, Abcam), t-STAT3 (1:400, Abcam), ROR-γt (1:400, Abcam) and GAPDH (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) antibodies at 4°C overnight. The anti-rabbit secondary antibody (1:10000, Li-Cor Bioscience, Bad Homburg, Germany) was incubated for 2 h at room temperature. The protein bands were detected by Odyssey Western Blot Detection System (LI-COR) and the band intensities were evaluated with ImageJ.

Liu X., Su Y., Liu J., Liu D, & Yu C. (2024). Inhibition of Th17 cell differentiation by aerobic exercise improves vasodilatation in diabetic mice. Clinical and experimental hypertension (New York, N.Y. : 1993), 46(1).

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