The largest database of trusted experimental protocols

Odyssey fluorescent scanner

Manufactured by LI COR
Sourced in United States

The Odyssey Fluorescent Scanner is a high-performance imaging system designed for detection and quantification of fluorescent signals in a variety of applications, including Western blotting, cell-based assays, and tissue imaging. The scanner utilizes a laser-based detection method to capture images with high sensitivity and resolution, allowing for accurate and reliable analysis of fluorescent samples.

Automatically generated - may contain errors

9 protocols using odyssey fluorescent scanner

1

Liver Protein Quantification and Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lysates were prepared from liver tissue and protein levels were determined by the bicinchoninic acid assay (Thermo Fisher, Carlsbad, CA). A total of 50 μg protein was applied to 10% SDS-PAGE gel, then transferred to PVDF membranes. The membranes were probed overnight at 4 °C with antibodies directed against PGC1α (#101707, Cayman Chemical, Ann Arbor, MI, 1:500) or GAPDH (MAB374, Millipore, Temecula, CA, 1:2000). After washing, membranes were probed with fluorescently-labeled secondary antibodies (Li-Cor, Lincoln, NE), and immunoreactive proteins detected using an Odyssey fluorescent scanner (Li-Cor, Lincoln, NE).
+ Open protocol
+ Expand
2

Quantifying PARP10 de-ADP-ribosylation Kinetics

Check if the same lab product or an alternative is used in the 5 most similar protocols
Following a previously established protocol (23 ), auto-mono-ADP-ribosylated PARP10cd was obtained by incubating 10 μM purified PARP10cd in 1 mM NAD+ for 20 min at 37˚C in Reaction Buffer (50 mM HEPES pH 8.0, 0.15 M NaCl, 0.2 mM TCEP, 0.02% NP-40). The de-ADP-ribosylation reaction was performed by incubating purified RuV macrodomain with mono-ADP-ribosylated PARP10cd at 37˚C in Reaction Buffer at equimolar ratios (1 μM). The reaction was terminated at 0, 1, 2, 4, 8, 16, 32 or 64 min by adding SDS-PAGE sample loading buffer and incubating at 95˚ for 5 min. Following separation SDS-PAGE on a 4-20% gradient gel, the proteins were transferred to a PVDF membrane and the de-MARylation activity determined by Western blotting. The membrane was blocked in 5% milk in PBS, 0.05% Tween-20. The membrane was incubated for 3 h at 22˚C in Anti-mono-ADP-Ribose Binding Reagent (Sigma, MABE1076, RRID:AB_2665469; 1:3,000 dilution) in place of a primary antibody. The secondary antibody was anti-rabbit immunoglobin (Cell Signaling Technology, 5151S, RRID:AB_10697505, DyLight® 800; dilution 1:10,000 dilution, 30 min at 22˚C). Blots were imaged with the near-infrared system of an Odyssey fluorescent scanner (LI-COR Biosciences) after washing with PBS and water.
+ Open protocol
+ Expand
3

Quantifying GRIP1 in Prefrontal Cortex

Check if the same lab product or an alternative is used in the 5 most similar protocols
GRIP1 levels in the prefrontal cortex were measured using a western blot, as described in Briand et al., 2014 (link). Briefly, animals were decapitated, and the prefrontal cortex dissected using a brain block (Braintree Scientific). Protein quantification was performed using a Pierce BCA Protein Assay Kit (Thermo Scientific). Equal amounts of protein (30 µg) were loaded into each well of a Tris-glycine gel (Lonza) and transferred to nitrocellulose membranes (Immobilon). Membranes were blocked with Li-Cor blocking buffer and allowed to incubate in primary antibody solution (GRIP1, 1:2000 (BD Biosciences) and GAPDH, 1:5000 (Cell Signaling)) for 24 hours at 4°C. Membranes were then incubated with fl uorescent secondary antibodies (1:20,000; IR-dye 680 or IR-dye 800, Li-Cor) and imaged on an Odyssey fluorescent scanner (Li-Cor). Western blots were normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and the percent knockout calculated as a fraction of the average of the GRIP1 levels in GFP-infused mice.
+ Open protocol
+ Expand
4

Western Blot Analysis of Hippocampal CRFR2

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole-cell hippocampal tissue was processed for Western blot as described previously (Anderson et al., 2008 (link)). Equal amounts of protein (20 μg) were loaded and separated in 10% Tris-Glycine gels (Invitrogen) and transferred to nitrocellulose membranes using the i-Blot dry transfer system (Invitrogen). Membranes were incubated overnight at 4°C with selective antibodies to CRFR2 (1:1000, Santa Cruz), or GAPDH (1:2000, Millipore). Membranes were then incubated with fluorescent secondary antibodies (1:5000, IR-dye 680 or IR-dye 800 Licor Biosciences), before being imaged on an Odyssey fluorescent scanner (Licor Biosciences). GAPDH was used as a loading control.
+ Open protocol
+ Expand
5

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were collected by scraping in 250 μL ice cold 1x RIPA buffer with protease inhibitor. Lysates were sonicated using a tip sonicator (Branson) followed by clearing of insoluble material by centrifugation. The protein concentration of the supernatants was estimated by absorbance measurements on a Nanodrop instrument. 30 μg of lysate per lane was resolved using 4–12% Bis-Tris SDS PAGE gels (Invitrogen) and then transferred to PVDF membrane (Bio-Rad). Membranes were blocked in 5% non-fat dry milk (Biorad) in TBST (Tris-buffered saline with 0.1% Tween 20) for 1 hr at room temperature. Primary antibodies were incubated in milk in TBST overnight at 4 °C. Primary antibodies used here were anti-TUBG1 (Sigma, T6557, 1:2000), FLAG (Sigma, M2, 1:2000), anti-NRF2 (Proteintech, 66504, 1:2000), anti-succinyllysine (PTM Biolabs, PTM-401, 1:1000), anti-MYC (and anti-SUCLG2 (Bethyl Laboratories, A305-533A, 1:2000). After 5x TBST washes for 15–30 minutes, fluorophore-conjugated (1:2000, LI-COR) or HRP-conjugated (1:3000, ThermoFisher) secondary antibodies were incubated in milk for 1 hr and then washed again with TBST before visualizing on a LI-COR Odyssey fluorescent scanner or with film.
+ Open protocol
+ Expand
6

Subcellular Protein Fractionation and Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were harvested with 2 mM EDTA in HBSS (Gibco) and washed twice with HBSS. Cell pellets were then lysed with Cell Lysis Buffer (CLB) (10 mM Hepes 7.6, 10 mM KCl, 1.5 mM MgCl2, 0.5% NP40, PMSF, protease/phosphatase inhibitors (HALT–Pierce). The cytosolic fraction was separated from the nuclear pellet. The nuclear pellet was lysed with Nuclear Lysis Buffer (NLB) (25 mM Tris 7.5, 1% SDS, protease/phosphatase inhibitors). Protein concentrations were determined using Pierce BCA Protein Assay.
Equivalent amounts of cytosolic or nuclear protein were loaded into wells of SDS–PAGE gels (4–12% Bis-Tris Gel, Thermo Fisher), and transferred to nitrocellulose. These membranes were incubated with the following antibodies at 4 °C overnight: Primary antibodies (Cell Signaling: ERK, phosphor-ERK, FGFR1-D8E4. Sigma: Actin). Secondary Antibodies (ThermoFisher: Goat anti Mouse Alexa Fluor 680, Goat anti Rabbit Alexa Fluor 800). Stained membranes were scanned with a LI-COR Odyssey Fluorescent Scanner.
+ Open protocol
+ Expand
7

Quantification of FoxO1 and Akt Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
WB analysis was used to quantify the expression of FoxO1, p-FoxO1, Akt, p-Akt, GPR17, and GAPDH active in signaling. After different treatments, total protein from SK-N-SH cells was extracted at 4 °C and the protein concentration quantified using a BCA protein assay kit. For each sample, 40 μg of protein was loaded. The following primary antibodies were used: anti-FoxO1 antibody (1:1000, Cell Signaling #2880), anti-p-FoxO1 Ser256 antibody (1:1000, Cell Signaling #9461), anti-Akt antibody (1:1000, Cell Signaling #4691), anti-p-Akt Ser473 antibody (1:1000, Cell Signaling #9271), anti-GPR17 antibody (1:500, Absin Bioscience Inc. #abs112096, Shanghai, China), and anti-GAPDH antibody (1:5000, Abcam #181602). The second antibody used was Goat anti-Rabbit IgG H&L (IRDye® 800 CW; 1:10,000, Abcam #216773). An Odyssey fluorescent scanner (LI-COR, Bioscience, Lincoln, NE, USA) was used to detect protein bands. The relative fold-change of protein expression levels was calculated by normalization to GAPDH levels using Quantity One software (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
+ Open protocol
+ Expand
8

Hippocampal Protein Analysis via Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mice were taken directly from their home cages and killed by cervical dislocation. Whole hippocampi were dissected and frozen in liquid nitrogen. Tissue was homogenized in 200 μL of ice-cold extraction buffer containing PBS, 1 mM EGTA, 1 mM EDTA, 0.01% SDS, and 1 mM PMSF. Protein concentration was quantified using the Pierce BCA Protein Assay Kit (Thermo Scientific). Equal amounts of protein (30 μg for whole cell) were loaded and separated in 10% polyacrylamide Tris–Glycine gels (Invitrogen) and transferred to nitrocellulose membranes using the i-Blot dry transfer system (Invitrogen). Membranes were blocked with Li-Cor blocking buffer. Membranes were incubated for 24 h at 4°C with selective antibodies to: CREB (1:1000; Santa Cruz), pCREB (1:1000, Millipore), TORC1 (1:1000, Cell Signaling), and GAPDH (1:2000, Cell Signaling). Membranes were then incubated with fluorescent secondary antibodies (1:5000, IR-dye 680 or IR-dye 800), before being imaged on an Odyssey fluorescent scanner (Licor Biosciences). GAPDH was used as an internal loading control.
+ Open protocol
+ Expand
9

Histone H3K9 methylation in B cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
B cells from four individuals with low KDM4C expression (GM11839, GM06994, GM12891, GM07034) and four individuals with high KDM4C expression (GM07056, GM12144, GM12815, GM12003) were washed in PBS and suspended in Triton lysis buffer (PBS with 0.5% Triton X 100 [v/v], 1× complete protease inhibitors [Roche], and 5 mM sodium butyrate) at 1 × 107 cells/mL and lysed for 10 min on ice. Cell nuclei were collected by centrifugation and resuspended in 250 μL of 0.2 M HCl and incubated overnight at 4°C to extract histones. The supernatant was collected, and 10–20 μg of protein were analyzed by Western blot with the following antibodies: 2 μg trimethylated H3K9 (17-625; Millipore), 2 μg dimethylated H3K9 (39683; Active Motif), 5 μg monomethylated H3K9 (39681; Active Motif), or 0.5 μg total H3 (Abcam). Quantitative Western blot detection was carried out using secondary antibodies coupled to fluorophores (Licor), and the fluorescent signal was detected on an Odyssey fluorescent scanner (Licor). A representative Western blot is shown in greyscale in Supplemental Figure S4. No significant difference in fluorescent signal was detected among individuals with high and low KDM4C expression for any of the methylation states of H3K9, by t-test. Histone extractions from B cells were repeated, and the same results were obtained.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!