Ir800 dye

Manufactured by LI COR

Sourced in United States

The IR800 dye is a near-infrared fluorescent dye designed for use in a variety of biological and biomedical applications. It has an excitation maximum at 785 nm and an emission maximum at 810 nm, making it suitable for detection in the near-infrared region of the spectrum.

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Lab products found in correlation

Anti tubulin, by Cell Signaling Technology (1 mentions) Anti lamin a c, by Santa Cruz Biotechnology (1 mentions) Anti gli1, by Cell Signaling Technology (1 mentions) Donkey anti goat hrp, by Jackson ImmunoResearch (1 mentions) Anti gli3, by R&D Systems (1 mentions) Donkey anti mouse hrp, by Jackson ImmunoResearch (1 mentions) Odyssey fc imaging system, by LI COR (1 mentions) Ir680, by LI COR (1 mentions) Bis tris gradient gel, by Thermo Fisher Scientific (1 mentions) Actin, by Merck Group (1 mentions) Fibronectin, by Abcam (1 mentions) Sds page, by Thermo Fisher Scientific (1 mentions) Cell lysis buffer, by Cell Signaling Technology (1 mentions) Blocking buffer, by LI COR (1 mentions) Alexa fluor 680, by LI COR (1 mentions) Phenylmethansulfonyl fluoride, by Merck Group (1 mentions) Odyssey system, by LI COR (1 mentions) β actin, by Merck Group (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Bradford assay, by Bio-Rad (1 mentions)

6 protocols using ir800 dye

Subcellular Fractionation and Western Blot Analysis

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Subcellular fractions were isolated using the Active Motif kit. Fractions were normalized for protein content, and analyzed by immunoblot. Blots were incubated in blocking buffer (5% non-fat dry milk, 1X TBS, 0.1% Tween) for 30 minutes, then incubated overnight at 4°C with anti-Gli3 (R&D), anti-Gli1 (Cell Signaling), anti-Lamin AC (Santa Cruz) and anti-tubulin (Cell Signaling) antibodies in blocking buffer. Blots were washed 3x in TBST buffer (1X TBS, 0.1% Tween) then incubated for 1 hour with donkey anti-mouse HRP (Jackson Immuno), donkey anti-goat HRP (Jackson Immuno), or IR 800 mouse (Li-COR). Blots were developed using an Odyssey Fc imaging system (Li-COR) and either ECL prime chemiluminescent substrate (GE Amersham) or IR800 dyes (Li-COR).

Arensdorf A.M., Dillard M.E., Menke J.M., Frank M.W., Rock C.O, & Ogden S.K. (2017). Sonic Hedgehog activates Phospholipase A2 to enhance Smoothened ciliary translocation. Cell reports, 19(10), 2074-2087.

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Western Blot Analysis of Cell Lysates

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Cells were harvested by lysis in the presence of protease inhibitors, and run on 4%–12% Bis-Tris gradient gel (Invitrogen). Transfer was done onto Nitrocellulose membrane and the membrane was blocked with 5% nonfat dried milk prior to antibody addition. p21 (1:1 000, Calbiochem), p53 (1:8 000, DO1 Santa Cruz), Actin (1:10 000, Sigma) and RFP (1:5 000, MBL) antibodies were used. Secondary antibodies with IR-680, IR-800 dyes (1:10 000, Licor) were used for detection.

Hafner A., Reyes J., Stewart-Ornstein J., Tsabar M., Jambhekar A, & Lahav G. (2020). Quantifying the Central Dogma in the p53 pathway in live single cells. Cell systems, 10(6), 495-505.e4.

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Fluorescent DNA Probe Synthesis

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Complementary oligonucleotides were synthesized with the top strand containing a 5’ C6 amino ester modification (Sigma). We mixed the top oligo with an equal molar ratio IR-800 dye (LiCOR) in Phosphate buffered saline (10 mM Na₂HPO₄, 1.8 mM KH₂PO₄, 137 mM NaCl, 2.7 mM KCl pH 7.4) and incubated for 6 hours protected from light. Next, we combined the labeled top strand with the complementary bottom strand and annealed them. Free dye and unannealed strands were removed by gel purification on a 4% Certified Low Range Ultra agarose gel (BioRad) in TAE buffer. The annealed strands were excised from the gel and extracted with a Nebulizer (Millipore) following the manufacturers protocol. The extracted probe was purified by ethanol precipitation and the DNA pellet was resuspended in EB buffer (10 mM Tris-HCl, pH 8.5).

Jackobel A.J., Zeberl B.J., Glover D.M., Fakhouri A.M, & Knutson B.A. (2019). DNA binding preferences of S. cerevisiae RNA polymerase I Core Factor reveal a preference for the GC-minor groove and a conserved binding mechanism. Biochimica et biophysica acta. Gene regulatory mechanisms, 1862(9), 194408.

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Protein Analysis from Aortic Tissue

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For protein analyses from whole aortic tissue, individual mouse aorta were powdered using a mortar and pestal and homogenized in Cell Lysis Buffer (Cell Signaling) containing 1 mM phenylmethansulfonylfluoride (Sigma). The lysates were then collected by pelleting the cell debris at 14,000 rpm for 10 minutes at 4°C. Total protein concentration was determined by Bradford Assay (BioRad). Protein lysates (50 ug per lane) were separated by 4–12% SDS PAGE (Invitrogen) and transferred onto a nitrocellulose membrane (BioRad). The membranes were blocked at room temperature with Blocking Buffer (LI-COR Biosciences) for 1 hour, incubated with appropriate antibodies (in LI-COR Blocking Buffer) overnight at 4°C, and washed three times with TBST. Secondary antibodies (1:10,000 in 50% Blocking Buffer/50% TBST) consisted of anti-rabbit and anti-mouse IgGs conjugated to Alexa-Fluor 680 and IR800Dye (LI-COR Biosciences). The blots were probed for Fibronectin (1:1,000; Abcam) and β-actin (1:5,000, Sigma) and were identified simultaneously (800 nm and 700 nm wavelengths, respectively) using near-infrared visualization (Odyssey System, LI-COR Biosciences). Densitometry was performed using the Odyssey software.

Chiasson V.L., Pakanati A.R., Hernandez M., Young K.J., Bounds K.R, & Mitchell B.M. (2017). REGULATORY T CELL AUGMENTATION OR INTERLEUKIN-17 INHIBITION PREVENTS CALCINEURIN INHIBITOR-INDUCED HYPERTENSION IN MICE. Hypertension (Dallas, Tex. : 1979), 70(1), 183-191.

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Antibody Binding to MSCs via PA-PG

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Cmab and an isotype control IgG were labeled with IR800 dye (LI-COR Biosciences, Lincoln, NE, USA) using established procedures. MSCs were incubated with PA-PG (50 µg/mL) in serum-free media for 1 h at 37 °C. Labeled Ab (either Cmab or isotype Ig) was added to PA-PG-coated MSCs and incubated for 1 h at 4 °C. Post-incubation, the cells were washed with DPBS, resuspended in flow buffer, and analyzed for IR800 fluorescence intensity via flow cytometry.

Nethi S.K., Li X., Bhatnagar S, & Prabha S. (2023). Enhancing Anticancer Efficacy of Chemotherapeutics Using Targeting Ligand-Functionalized Synthetic Antigen Receptor-Mesenchymal Stem Cells. Pharmaceutics, 15(6), 1742.

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VSX-1 Conjugation and In Vivo Imaging

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CR female NCI Ath/nu mice were placed on a special "RD D10012Mi" diet for 7 days prior to the start of the study and for the duration of the study. The animals were randomized into treatment groups based on the Day 1 bodyweight, with an age at start date of 8 to 12 weeks. VSX was conjugated with IR800 dye (LiCor, Lincoln, NE) and then used in sortase reactions to make VSX-1. A vehicle control (0.9% saline) and a dye-only control were also employed. A 5 mg/kg dose was used, and substances were injected IV. Whole-body imaging (dorsal and ventral) was performed at 1, 6, 12, 24, 48 and 100 hours post-IV dosing.

Johnson K., Delaney J.C., Guillard T., Reffuveille F., Varin-Simon J., Li K., Wollacott A., Frapy E., Mong S., Tissire H., Viswanathan K., Touti F., Babcock G.J., Shriver Z., Pentelute B.L., Plante O, & Skurnik D. (2023). Development of an antibody fused with an antimicrobial peptide targeting Pseudomonas aeruginosa: A new approach to prevent and treat bacterial infections. PLOS Pathogens, 19(9), e1011612.

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