Texas red x succinimidyl ester

Manufactured by Thermo Fisher Scientific

Texas Red-X succinimidyl ester is a fluorescent dye used in various biomedical applications. It is a reactive ester derivative of the Texas Red fluorescent dye, which can be used to label proteins, peptides, and other biomolecules. The succinimidyl ester group allows for covalent attachment of the dye to amine-containing molecules.

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Lab products found in correlation

Alexa fluor 488 carboxylic acid, by Thermo Fisher Scientific (1 mentions) Lysotracker blue dnd 22, by Thermo Fisher Scientific (1 mentions) Calcium assay kit, by BioAssay Systems (1 mentions) Collagenase, by Thermo Fisher Scientific (1 mentions) N hydroxysulfosuccinimide, by Merck Group (1 mentions) Quant it picogreen dna assay kit, by Thermo Fisher Scientific (1 mentions) 5 aminomethyl fluorescein hydrochloride, by Thermo Fisher Scientific (1 mentions) Human bmp2 elisa kit, by Merck Group (1 mentions) Human bmp 2 protein, by Medtronic (1 mentions) Sodium β glycerophosphate, by Merck Group (1 mentions) Rat tail collagen type 1, by Corning (1 mentions) Ascorbic acid, by Merck Group (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) N 3 dimethylaminopropyl n ethylcarbodiimide hydrochloride, by Merck Group (1 mentions)

9 protocols using texas red x succinimidyl ester

Mouse Myocardial Infarction Model

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Example 7

Mouse Model of Myocardial Infarction:

The method to induce myocardial infarction in mice was based on previous studies (Andrade et al., (2015) PLoS One 10: e0143221). Briefly, male CD1 mice were anesthetized with 3% isofluorane combined with 2% oxygen inhalation. Under sterile conditions, the heart was exposed by a minimally invasive left thoracotomy and acute myocardial infarction (AMI) was produced by permanent ligation of the left anterior descending coronary artery. Immediately after AMI induction, the heart was randomized to receive one of the following four treatment arms: 1) “Control (PBS)” group: Intramyocardial injection of 50 μl PBS into the heart immediately after AMI; 2) “Control MP₁” group: Intramyocardial injection of 1×10⁵Control MP₁in 50 μl PBS into the heart immediately after AMI; 3) “CMMP” group: Intramyocardial injection of 1×10⁵CMMPs in 50 μl PBS into the heart immediately after AMI; 4) “CSC” group: Intramyocardial injection of 1×10⁵CSCs in 50 μl PBS into the heart immediately after AMI. To enable visualization of Control MP₁or CMMP in a cohort of animals, the Control MP₁or CMMP were pre-labeled with Texas Red-X succinimidyl ester (1 mg/ml [Invitrogen, Carlsbad, Calif.]).

US11564889B2. Stem cell biomimetic microparticles (2023-01-31). North Carolina State University [US]. Inventors: Ke Cheng [US], Junnan Tang [US].

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Gentamicin-Texas Red Conjugation Protocol

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Gentamicin–Texas Red conjugate (Gent: TR) was prepared as previously described (Dai et al., 2006 (link)) with slight modifications. Briefly, 440 μl of 50 mg gentamicin sulfate ml^–1 was mixed with 60 μl of 2 mg amine-reactive Texas Red-X succinimidyl ester (Invitrogen) ml^–1 in anhydrous N, N-dimethylformamide. The mixture was gently rotated for 3 days at 4°C to produce an approximately 30: 1 molar ratio (∼10 mM gentamicin and 0.3 mM Texas Red reagent) of Gen: TR. Next, 100 μl of the Gen: TR was added to the stationary phase cells (2 × 10⁷), washed, and resuspended in 2 ml PBS for 3 h at 37°C with continuous rotation. After incubation, the cells were washed twice, resuspended in 500 μl PBS, and evaluated by flow cytometry with excitation/emission maxima at 595/615 nm.

Pandey S., Sahukhal G.S, & Elasri M.O. (2021). The msaABCR Operon Regulates Persister Formation by Modulating Energy Metabolism in Staphylococcus aureus. Frontiers in Microbiology, 12, 657753.

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Synthesis of Fluorescent Gelatin

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Fluorescent gelatin was synthesized by incubating bovine skin gelatin with amine reactive fluorescent dyes. Alexa Fluor 488 carboxylic acid, 2,3,5,6-tetrafluorophenyl ester (Invitrogen), or Texas red-X succinimidyl ester (Invitrogen) was dissolved in DMSO at 10 mg/ml. Dye solution (50 µl) was added slowly to 1 ml of bovine skin gelatin solution (Sigma-Aldrich; 40 mg/ml; dissolved in 0.1 M sodium bicarbonate buffer, at pH 8.3 for succinimidyl ester or pH 9.0 for tetrafluorophenyl ester) while vortexing. After incubation at room temperature for 1 h, the free dyes in the mixture were removed with a HiTrap desalting column (GE Healthcare). The desalted fluorescent gelatin solution can be stored at 4°C for short-term storage or at −80°C with 10% glycerol as a cryoprotectant for long-term storage.

Sun J., Lu F., He H., Shen J., Messina J., Mathew R., Wang D., Sarnaik A.A., Chang W.C., Kim M., Cheng H, & Yang S. (2014). STIM1- and Orai1-mediated Ca2+ oscillation orchestrates invadopodium formation and melanoma invasion. The Journal of Cell Biology, 207(4), 535-548.

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Murine Myocardial Infarction Treatment

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All animal
work was approved by the Institutional Animal Care and Use Committee
at North Carolina State University. Mouse MI model was generated as
previously described.^{45 (link),46 (link)} Generally, male CD1 mice were
anesthetized with isoflurane mixed with oxygen inhalation. The heart
was exposed by a minimally invasive left thoracotomy, and LAD was
ligated permanently for induction of acute MI. After LAD ligation,
the heart was to receive one of the following four treatments randomly:
(1) MI + hCSCs in nanogel, intramyocardially injected with 1 ×
10⁵ hCSCs in 50 μL of P(NIPAM-AA) nanogel; (2) MI
+ hCSCs in PBS, intramyocardially injected with 1 × 10⁵ hCSCs in 50 μL of PBS; (3) MI + nanogel, intramyocardially
injected with 50 μL of P(NIPAM-AA) nanogel; (4) MI alone, MI
surgery without any injection. The hCSCs or nanogels were prelabeled
with Texas Red-X succinimidyl ester (1 mg/mL, Invitrogen) for detection.

Tang J., Cui X., Caranasos T.G., Hensley M.T., Vandergriff A.C., Hartanto Y., Shen D., Zhang H., Zhang J, & Cheng K. (2017). Heart Repair Using Nanogel-Encapsulated Human Cardiac Stem Cells in Mice and Pigs with Myocardial Infarction. ACS Nano, 11(10), 9738-9749.

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Mouse Myocardial Infarction Treatment Protocol

Cited 1 time

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All animal work was compliant with the Institutional Animal Care and Use Committee at North Carolina State University. The method to induce myocardial infarction in mice was based on previous studies30 (link). Briefly, male SCID Beige mice were anaesthetized with 3% isofluorane combined with 2% oxygen inhalation. Under sterile conditions, the heart was exposed by a minimally invasive left thoracotomy and acute myocardial infarction (AMI) was produced by permanent ligation of the LAD coronary artery. Immediately after AMI induction, the heart was randomized to receive one of the following four treatment arms: (1) ‘Control (PBS)’ group: intramyocardial injection of 50 μl PBS into the heart immediately after AMI; (2) ‘Control MP₁’ group: intramyocardial injection of 1 × 10⁵ Control MP₁ in 50 μl PBS into the heart immediately after AMI; (3) ‘CMMP’ group: intramyocardial injection of 1 × 10⁵ CMMPs in 50 μl PBS into the heart immediately after AMI; (4) ‘CSC’ group: intramyocardial injection of 1 × 10⁵ CSCs in 50 μl PBS into the heart immediately after AMI. To enable visualization of Control MP₁ or CMMP in a cohort of animals, we pre-labelled the Control MP₁ or CMMP with Texas Red-X succinimidyl ester (1 mg ml⁻¹ (Invitrogen, Carlsbad, California)).

Tang J., Shen D., Caranasos T.G., Wang Z., Vandergriff A.C., Allen T.A., Hensley M.T., Dinh P.U., Cores J., Li T.S., Zhang J., Kan Q, & Cheng K. (2017). Therapeutic microparticles functionalized with biomimetic cardiac stem cell membranes and secretome. Nature Communications, 8, 13724.

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Myocardial Infarction Therapy with Microgels

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Male CD1 mice were anaesthetized with isoflurane-oxygen inhalation. Under sterile conditions, the heart was exposed through a minimally invasive left thoracotomy. MI was produced via the permanent ligation of the LAD coronary artery. Immediately after MI induction, the heart was randomized to receive one of the following three treatments: (1) “control (PBS)” group: intramyocardial injection of 50 μL of PBS; (2) “microgel” group: intramyocardial injection of P(NIPAM-IA) microgels; (3) microgel + hCSC group: intramyocardial injection of microgel-encapsulated hCSCs. The MI mice treated with microgel-encapsulated hCSCs were intraperitoneally injected with rapamycin. Texas Red-X succinimidyl ester (Invitrogen, Carlsbad) at 1 mg/mL was used to pre-label hCSCs and/or microgels for detection.

Cui X., Tang J., Hartanto Y., Zhang J., Bi J., Dai S., Qiao S.Z., Cheng K, & Zhang H. (2018). NIPAM-based Microgel Microenvironment Regulates the Therapeutic Function of Cardiac Stromal Cells. ACS applied materials & interfaces, 10(44), 37783-37796.

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Fluorescent Labeling and Imaging of LtxA

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Purified LtxA was fluorescently labeled with Texas Red-X Succinimidyl Ester (Thermo Fisher Science) in the following manner. LtxA was hydrated in PBS at a concentration of 0.125 μg/μL. The dye, dissolved in DMSO at 50 mg/mL, was mixed with the toxin at a concentration of 1.0 μg dye/μL protein. The mixture was shaken gently for one hour in the dark, then dialyzed overnight against 4 L of PBS at 4°C in the dark.
K562 cells (Lozzio and Lozzio, 1975 (link)) expressing a cyan fluorescent protein (CFP)-tagged αL cytosolic domain and a yellow fluorescent protein (YFP)-tagged β2 domain (Kim et al., 2003 (link)) at a concentration of 0.6 × 10⁶ cells/mL were labeled with Lysotracker® blue DND-22 (Thermo Fisher Scientific) for 30 min at 37 °C and then washed. Texas Red-labeled LtxA (TR-LtxA) was added at a concentration of 1 × 10⁻⁸ M; the cells were incubated for 30 min at 37 °C and then washed twice in PBS. In a second experiment, the cells were first incubated with 5 μM DRAQ5^™ for 30 mins, followed by Lysotracker® blue and TR-LtxA.

Webb J.N., Koufos E, & Brown A.C. (2016). Inhibition of Bacterial Toxin Activity by the Nuclear Stain, DRAQ5™. The Journal of membrane biology, 249(4), 503-511.

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Collagen-Alginate Hydrogel for Bone Tissue Engineering

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Rat tail collagen type I was purchased from Corning Inc. (Corning. NY, USA). Sodium alginate (alginate, 500GM) was purchased from Pfaltz & Bauer Inc. (Waterbury, CT, USA). Sodium hydroxide (NaOH), ethanol, 5-(Aminomethyl)Fluorescein Hydrochloride, and Texas Red™-X, Succinimidyl Ester were purchased from Thermo Fisher Scientific (Waltham, MA). Calcium sulfate (CaSO₄), 2-(N-Morpholino)ethanesulfonic acid (MES), N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC), N-Hydroxysulfosuccinimide (NHS), dexamethasone, ascorbic acid, β-sodium glycerophosphate, and Human BMP2 ELISA kit were received from Sigma-Aldrich (St Louis, MO). Collagenase, Calcein AM/Ethidium homodimer-1 Live/Dead assay kit, and Quant-iT PicoGreen DNA assay kit were purchased from Thermo Fisher Scientific (Waltham, MA). QuantiChrom alkaline phosphatase activity (ALP) kit and Calcium Assay kit were purchased from BioAssay Systems (Hayward, CA, USA). Human BMP2 protein was provided by Medtronic (Dublin, Ireland).

Moeinzadeh S., Park Y., Lin S, & Yang Y.P. (2020). In-situ stable injectable collagen-based hydrogels for cell and growth factor delivery. Materialia, 15, 100954.

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Aminoglycoside Uptake Assay Protocol

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The aminoglycoside uptake assay was done according to Loughman et al. (2016) (link). GEN was tagged using the Texas Red-X succinimidyl ester (ThermoFisher) at a molar ratio of 300:1 and incubated overnight at 4°C with agitation. To remove unbound Texas Red, the dye remover column (Thermo Fisher) was used according to the manufacturer’s protocol. Bacteria were adjusted to an A₆₀₀_nm of 0.1 and incubated 2 h at 35°C with agitation in the presence of 128 μg/ml of the Texas Red-GEN conjugate. When TO was added during the uptake assay, it was used at 8 μg/ml. Plates were read with a TECAN Genios Fluorescence plate reader with the 535/595 nm filters.

Langlois J.P., Millette G., Guay I., Dubé-Duquette A., Chamberland S., Jacques P.É., Rodrigue S., Bouarab K., Marsault É, & Malouin F. (2020). Bactericidal Activity of the Bacterial ATP Synthase Inhibitor Tomatidine and the Combination of Tomatidine and Aminoglycoside Against Persistent and Virulent Forms of Staphylococcus aureus. Frontiers in Microbiology, 11, 805.

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