Odyssey gel imager

Manufactured by LI COR

The Odyssey gel imager is a fluorescence imaging system designed for the detection and quantification of proteins and nucleic acids separated by gel electrophoresis. It uses high-sensitivity fluorescence detection to capture images of stained gels or blots. The Odyssey system is capable of scanning a variety of gel and blot formats.

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Lab products found in correlation

Quantstudio 3 real time pcr system, by Thermo Fisher Scientific (2 mentions) Iblot system, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by LI COR (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Ab76020, by Abcam (1 mentions) Anti rabbit secondary antibody, by LI COR (1 mentions) Hpa010856, by Merck Group (1 mentions) Immobilon fl, by Merck Group (1 mentions) Biotinylated antibodies, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Phagocytosis assay kit, by Cayman Chemical (1 mentions) Spot digital camera, by Nikon (1 mentions) Cd11b magnetic separation kit, by STEMCELL (1 mentions) Luminex multiplex cytokine assay, by Thermo Fisher Scientific (1 mentions) Dmem f12, by Thermo Fisher Scientific (1 mentions) Celltiter glo luminescent cell viability assay kit, by Promega (1 mentions) Cell culture supplies, by Corning (1 mentions)

Close spelling variants of this product

Odyssey infrared gel imager Odyssey Imager

5 protocols using odyssey gel imager

Quantitative Analysis of Intracellular Aggregates

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Western blots were quantified by direct measurements of the secondary antibody fluorescence using Odyssey gel imager (LiCor, Lincoln, NE). Quantification of the band intensity over the background was calculated using the Odyssey gel imaging software integrated with the instrument.
For description of intracellular S129-positive aggregates, the following parameters were measured:

“total intensity” (following the command name in the Metamorph imaging software) measures the total gray level of each aggregate, thresholded against the background; this parameter directly depends on the area and density of the aggregate and thus quantitatively reflects the aggregate size.

“average intensity” (following the command name in the Metamorph imaging software) measures the gray levels (equaling fluorescence intensity) per unit area, and this reflects the density of the aggregate, as more densely packed aggregate would have higher fluorescence per unit area.

Zhao J., Pan B., Fina M., Huang Y., Shimogawa M., Luk K.C., Rhoades E., Petersson E.J., Dong D.W, & Kashina A. (2022). α-Synuclein arginylation in the human brain. Translational Neurodegeneration, 11, 20.

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Tyrosine Phosphorylation of Draper Domains

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To determine the degree of Tyr phosphorylation on His₁₀-Draper intracellular domains, liposome phosphorylation assays were performed as above, quenched using SDS-PAGE buffer + DTT, and incubated for 5 min at 95°C. Reactions were run on 4–20% gradient SDS-PAGE gels and transferred onto nitrocellulose membranes using the iBlot system (Invitrogen). Membranes were blocked for 1 h at room temperature in PBS + 0.1% wt/vol BSA (Sigma-Aldrich) and incubated overnight at 4°C with anti-His probe (rabbit) and anti-pTyr (mouse) antibodies (Table S1). Primary antibodies were incubated with membranes simultaneously. Both were used at 1:1,000 final dilution PBS + 0.1% wt/vol BSA. Membranes were washed 3 × 5 min at room temperature in PBS + 0.1% wt/vol BSA and incubated with Li-Cor secondary antibodies at 1:10,000 dilution (700 nm anti-rabbit and 800 nm anti-mouse; Table S1) for 1 h at room temperature. Membranes were washed 3 × 5 min at room temperature in PBS + 0.1% wt/vol BSA and imaged using a Li-Cor Odyssey gel imager. Quantification of anti-pTyr and anti-His intensity was performed using the Gel Analysis feature of Fiji (ImageJ; National Institutes of Health).

Williamson A.P, & Vale R.D. (2018). Spatial control of Draper receptor signaling initiates apoptotic cell engulfment. The Journal of Cell Biology, 217(11), 3977-3992.

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Na/K ATPase and FXYD3 Protein Analysis

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Samples were separated by SDS-PAGE and transferred to PVDF membranes (Immobilon-FL, Millipore). The total protein for each lane was detected with revert total protein stain and washed as per manufacturer’s protocol (LI-COR). Following total protein detection, the membrane was blocked with 0.1% casein protein in PBS for 1 h at room temperature. Then, the membrane was concurrently probed with rabbit Na/K ATPase (1:1,000 dilution; Abcam; ab76020; Lot GR3184452‐8) and rabbit FXYD3 (1:250 dilution, Sigma-Aldrich; HPA010856; Lot A57803) antibodies for 2 h at room temperature. Following PBS washes, the membrane was incubated with a 1:10,000 dilution of anti-rabbit secondary antibody (LI-COR). After a final PBS wash, images were obtained with an Odyssey gel imager (LI-COR), and raw signal intensities were analyzed with ImageJ (v2.3.0/1.53f, NIH) using the approach of the NIH ImageJ user guide. We used the area under both bands observed with the Na/K ATPase for analysis because heating the sample induces Na/K ATPase dimers (11 (link)). Total protein staining was the gel loading control, and data are presented as the %FXYD3 siRNA normalized to its donor-matched siRNA control.

Cano Portillo C., Villacreses R., Thurman A.L., Pezzulo A.A., Zabner J, & Thornell I.M. (2022). FXYD3 facilitates Na+ and liquid absorption across human airway epithelia by increasing the transport capacity of the Na/K ATPase. American Journal of Physiology - Cell Physiology, 323(4), C1044-C1051.

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Multimodal Characterization of Macrophage Response

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Dulbecco’s modified Eagle’s medium (DMEM), DMEM-F12, fetal bovine serum (FBS), L-glutamine (Q), penicillin/streptomycin (P/S), Sodium Pyruvate (SP), and Non-Essential Amino Acids (NEAA) were obtained from Invitrogen (Carlsbad, CA). CellTiter Glo Luminescent Cell Viability Assay kit was obtained from Promega (Madison, WI). The CD11b magnetic separation kit was purchased from Stem Cell Technologies (Vancouver, Canada). All standards used for the Luminex multiplex cytokine assay were purchased from PeproTech, Inc (Rocky Hill, NJ). Streptavidin-Biotin and biotinylated antibodies used for Luminex were purchased from eBioSciences (San Diego, CA). Phagocytosis assay kit was purchased from Cayman Chemicals (Ann Arbor, MI).
For qRT-PCR, the QuantStudio3 Real Time PCR system from Thermo Fisher (Waltham, MA) was used. For MTS and Griess assays, a Molecular Devices (Sunnyvale, CA) plate reader was used. For scanning Western blot, a Li-COR Odyssey gel imager (Lincoln, NE) was used. Cell culture supplies were purchased from Corning (Corning, NY). For immunocytochemistry studies, the images were obtained using a Nikon (Model: TE-2000U) inverted fluorescent microscope and images were obtained using a Spot Digital Camera (Sterling Heights, MI).

Sarkar S., Malovic E., Sarda D., Lawana V., Rokad D., Jin H., Anantharam V., Kanthasamy A, & Kanthasamy A.G. (2018). Characterization and comparative analysis of a new mouse microglial cell model for studying neuroinflammatory mechanisms during neurotoxic insults. Neurotoxicology, 67, 129-140.

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Western Blot Analysis of Signaling Proteins

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Wells were washed x2 with cold PBS and harvested in RiPA buffer containing total protease and phosphatase inhibitors. We quantified total protein by BCA assay and prepared samples at 10 μg/lane by resuspending in Laemmli buffer and heating for 10 min. at 70°C. Total protein samples were separated by SDS-PAGE using a BOLT 4-12% 1 mm 10-well gel and MES running buffer. We transferred samples onto a 0.45 μm PVDF membrane overnight at 4°C by wet transfer with Towbin buffer (containing 0.025% SDS and 10% MeOH). The membrane was blocked with 5% BSA in PBS-T (0.1% Tween-20) for 1 hr. at room temperature. Following, the membrane was incubated overnight at 4°C in primary antibodies diluted in blocking buffer at the following concentrations: TSC1 (1:1000), TSC2 (1:5000), ACTB (1:5000), pS6RP Ser235/236 (1:5000), S6RP (1:500), p4E-BP1 Ser65 (1:1000), 4E-BP1 (1:1000), pS6K1 Thr389 (1:500), S6K1 (1:1000), and HIF1A (1:500). The membrane was washed 3 × 5 min. with PBS-T, and then incubated for 1 hr. at room temperature in fluorescent secondary antibodies diluted 1:10,000 in blocking buffer. The membrane was washed again 3 × 5 min. with PBS-T and then imaged using Odyssey Gel Imager (LI-COR Biosciences). Samples were analyzed in ImageJ by quantifying signal intensity, normalizing by unphosphorylated protein or ACTB, and scaling between 0 to 1.

Pietrobon A., Yockell-Lelièvre J., Flood T.A, & Stanford W.L. (2022). Renal organoid modeling of tuberous sclerosis complex reveals lesion features arise from diverse developmental processes. Cell reports, 40(1).

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