Alexa fluor 594 nhs ester

Manufactured by Thermo Fisher Scientific

Sourced in United States

Alexa Fluor 594 NHS Ester is a fluorescent dye used in various biological applications. It is a succinimidyl ester derivative of the Alexa Fluor 594 dye, which can be used to label proteins and other biomolecules. The NHS ester group allows the dye to form covalent bonds with primary amines, enabling the labeling of amine-containing compounds.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Alexa Fluor 594 (2045 citations) Alexa 594 (538 citations) Alexa Fluors (21 citations) Alexa 594 NHS (2 citations) NHS-esters (2 citations)

20 protocols using alexa fluor 594 nhs ester

Immunohistochemical Profiling of Muscle Fiber Types

Check if the same lab product or an alternative is used in the 5 most similar protocols

Gastrocnemius muscles were freshly frozen in isopentane cooled in liquid nitrogen, after which transverse 20-μm cryostat sections were used for muscle immunohistochemistry and morphological analysis. After mounting on MAS-coated slides (Matsunami glass), the sections were fixed in cold acetone and processed for immunohistochemical staining as described above using mouse monoclonal antibodies to the type I, IIA and IIB myosin heavy chain (MyHC) isoforms (HB287, HB277 and HB283, respectively; ATCC). Hybridoma clones HB287 and HB277 were intraperitoneally injected into nude mice, and the ascites fluid was harvested and labeled with Alexa Fluor594 NHS Ester (Molecular Probes). For HB283, the IgM fraction was purified from culture supernatant using HiTrap IgM Purification HP (GE), and Alexa Fluor594-labeled donkey anti-mouse IgM, μ Chain Specific (Jackson ImmunoResearch Laboratories) was used as a secondary antibody for detection. For morphological analysis, cryosections were stained with hematoxylin and eosin.

Morisaki Y., Niikura M., Watanabe M., Onishi K., Tanabe S., Moriwaki Y., Okuda T., Ohara S., Murayama S., Takao M., Uchida S., Yamanaka K, & Misawa H. (2016). Selective Expression of Osteopontin in ALS-resistant Motor Neurons is a Critical Determinant of Late Phase Neurodegeneration Mediated by Matrix Metalloproteinase-9. Scientific Reports, 6, 27354.

+ Open protocol

+ Expand

Fabrication of MSC-Sec-loaded HA Hydrogel

Check if the same lab product or an alternative is used in the 5 most similar protocols

MSC-Sec-loaded, crosslinked HA gel was fabricated as follows: Briefly, 0.2 g of HA (Sigma-Aldrich, MO, USA), in a sodium salt state, was dissolved in 20 mL of ultrapure water and stirred at 200 rpm for 20 min. Then, 0.8 mL of a 5 n sodium hydroxide (Sigma-Aldrich) solution was added and reacted with 1 mL of methacrylic anhydride (Sigma-Aldrich). The resultant solution was stirred at 400 rpm for 2 h. It was then stored at 4 °C for 24 h, after which it was mixed with 95% ethanol to precipitate. Then, the sediments were washed with 95% ethanol, placed in 40 °C for 3 h, dissolved in ultrapure water, frozen thoroughly, and finally lyophilized. The powder was dissolved by ultrapure water and dialyzed, using a 12 kDa cellulose bag (Spectrum Laboratories, Rancho Dominguez, CA) immersed in 1 L ultrapure water, which served as buffer medium. After 1 week, the resultant MA-HA was lyophilized. The lyophilized Sec was combined with the MA-HA solution and exposed to UV light at a distance of 1.5 cm, using a wavelength of 365 nm (BlueWave 75 UV Curing Spot Lamp), at a power of 4.5 mW cm⁻², for 10 s. The final product was the MSC-Sec-loaded, crosslinked hyaluronic acid gel. MA-HA was reacted with cysteamine first to produce amino groups, and then the Alexa Fluor 594 NHS Ester (A20004, Molecular Probes) was used to react with amino groups to fluorescently label MA-HA.

Liu F., Hu S., Yang H., Li Z., Huang K., Su T., Wang S, & Cheng K. (2019). Hyaluronic Acid Hydrogel Integrated with Mesenchymal Stem Cell-Secretome to Treat Endometrial Injury in a Rat Model of Asherman’s Syndrome. Advanced healthcare materials, 8(14), e1900411.

+ Open protocol

+ Expand

Protein Corona Mediated Nanoparticle Uptake

Check if the same lab product or an alternative is used in the 5 most similar protocols

After incubation in plasma under the different flow conditions and washing in PBS to remove the soft corona, the PC coated PS-NPs were fluorescently labeled with Alexa Fluor 594 NHS ester (ThermoFisher Scientific, Waltham, MA). The NPs were sonicated for 5 minutes in a bath sonicator (VWR® Ultrasonic Cleaners) to break down the aggregates before addition to cells.
The protein corona coated NPs (25 μg mL⁻¹) were incubated with 4T1 cells at a seeding density of 10 000 cells per well in a 96 well-plate, in serum-free media for 4 hours. The cells were washed three times with phosphate-buffered saline (PBS), stained with Hoechst 33258 dye, and fixed in 4% paraformaldehyde before imaging.

Conjeevaram S.B., Blanchard R.M., Kadaba A, & Adjei I.M. (2022). Vascular bifurcation influences the protein corona composition on nanoparticles and impacts their cellular uptake. Nanoscale Advances, 4(12), 2671-2681.

+ Open protocol

+ Expand

Labeling of Enterovirus A71 with Alexa Fluor™ 594

Check if the same lab product or an alternative is used in the 5 most similar protocols

EVA71 was concentrated by ultracentrifugation at 120,000 g for 4 h at 4 °C (SW32Ti rotor, Beckman Coulter Optima L-100XP). The viral titer of the concentrated EVA71 was measured by plaque assay. Then 3 × 10⁸ PFU of EVA71 was labeled with Alexa Fluor™ 594 NHS Ester (A37572, ThermoFisher) following the instruction of the manuscript. The labeled EVA71 was aliquoted and stored at −80 °C, and the titer was measured by plaque assay.

Yan J., Wang M., Wang M., Dun Y., Zhu L., Yi Z, & Zhang S. (2020). Involvement of VCP/UFD1/Nucleolin in the viral entry of Enterovirus A species. Virus Research, 283, 197974.

+ Open protocol

+ Expand

Quantifying Cellular Uptake of Fluorescent Probes

Check if the same lab product or an alternative is used in the 5 most similar protocols

CLC or BSA (10735078001; Sigma‐Aldrich) was labeled with Alexa Fluor™ 488 NHS Ester (Thermo Fisher Scientific) or Alexa Fluor™ 594 NHS Ester (Thermo Fisher Scientific) and incubated with 4T1 cells for 2 h at 37°C. After five washes with PBS buffer, 4T1 cells were stained with lysosomal staining kit (ab112137; Abcam). 4T1 cells were fixed with 4% paraformaldehyde (Electron Microscopy Sciences), and diamidino‐2‐phenylindole (DAPI) (VECTASHIELD, Vector Laboratories) was used to counterstain the cell nuclei. The cells were visualized using an EVOS™ FL Auto 2 Imaging System (Thermo Fisher Scientific). After splitting each color channel, the area with fluorescence was measured for each color channel with Image J (National Institutes of Health; Bethesda, MD). The percent of uptake of CLC was calculated by dividing with measured area from DAPI channel.

Jung S., Jiang L., Zhao J., Shultz L.D., Greiner D.L., Bae M., Li X., Ordikhani F., Kuai R., Joseph J., Kasinath V., Elmaleh D.R, & Abdi R. (2022). Clathrin light chain‐conjugated drug delivery for cancer. Bioengineering & Translational Medicine, 8(1), e10273.

+ Open protocol

+ Expand

Preparation of Fluorescent Aspergillus fumigatus Conidia

Check if the same lab product or an alternative is used in the 5 most similar protocols

The A. fumigatus strain AfS150 [22 (link)], generated from ATCC 46645 and expressing the dTomato fluorescent protein, was used in this study. The fungus was grown at 37°C on Aspergillus minimal medium supplemented with 1% D-glucose as the carbon source. A fungal suspension was transferred to AMM agar plates and incubated for 3 days at 37°C. Conidia were harvested in 0.01% Tween 20–Dulbecco's Phosphate-Buffered Saline (DPBS) (PanEco, Russia) solution.
Conidia were then fixed overnight with 3% paraformaldehyde (Sigma-Aldrich, USA), washed twice with DPBS, filtered through Steriflip Filter Units (Millipore, Ireland), aliquoted, and stored at 4°C until use. Since the fluorescence of dTomato fluorescent protein was lost after fixation, they were labeled with Alexa Fluor 594 NHS Ester (Thermo Fisher, USA, A20004) for visualization, according to the manufacturer's instructions. Alexa Fluor 594-labeled spores were then filtered through Steriflip Filter Units (Millipore, Ireland), aliquoted, and stored at 4°C until use.

Shevchenko M.A., Bogorodskiy A.O., Troyanova N.I., Servuli E.A., Bolkhovitina E.L., Büldt G., Fahlke C., Gordeliy V.I., Gensch T., Borshchevskiy V.I, & Sapozhnikov A.M. (2018). Aspergillus fumigatus Infection-Induced Neutrophil Recruitment and Location in the Conducting Airway of Immunocompetent, Neutropenic, and Immunosuppressed Mice. Journal of Immunology Research, 2018, 5379085.

+ Open protocol

+ Expand

Fluorescent BSA-PEG Droplet Preparation

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

BSA-PEG droplets were prepared as
previously described.^{32 (link)} Briefly, we formed
the droplets by preparing a solution of 100 mM potassium phosphate,
pH 7, (Acros Organics, 42420 and Merck, 60349), 200 mM KCl (Merck,
60128), 30 g/L bovine serum albumin (BSA) (Merck, A7638), and 230
g/L PEG 4000 (Alfa Aesar, A16151) in Milli-Q water. In order to visualize
the droplets with fluorescence imaging, a small fraction of the BSA
was fluorescently labeled using Alexa Fluor594 NHS ester (ThermoFisher,
A20004). This resulted in droplets enriched in BSA suspended in a
supernatant enriched in PEG, both containing phosphate buffer and
salt.

Testa A., Spanke H.T., Jambon-Puillet E., Yasir M., Feng Y., Küffner A.M., Arosio P., Dufresne E.R., Style R.W, & Rebane A.A. (2023). Surface Passivation Method for the Super-repellence of Aqueous Macromolecular Condensates. Langmuir, 39(41), 14626-14637.

+ Open protocol

+ Expand

Fluorescent Labeling of rDGCR8 and rDROSHA

Check if the same lab product or an alternative is used in the 5 most similar protocols

The above rDGCR8 protein was labeled with Alexa Fluor 594 by using Alexa Fluor™ 594 NHS Ester (Succinimidyl Ester, ThermoFisher), and the rDROSHA protein was labeled with Alexa Fluor 488 by using Alexa Fluor™ 488 NHS Ester (Succinimidyl Ester, ThermoFisher) according to the manufacturer’s instructions. Briefly, recombinant proteins were incubated with the protein reaction buffer (20 mM HEPES pH 8.3, 150 mM NaCl and 1 mM DTT), and rotated at room temperature for 1 h. Free dye was removed by gel-filtration.

Cui Y., Lv X., Ding L., Ke L., Yang D., Pirouz M., Qi Y., Ong J., Gao G., Du P, & Gregory R.I. (2021). Global miRNA dosage control of embryonic germ layer specification. Nature, 593(7860), 602-606.

+ Open protocol

+ Expand

Fluorescent Labeling of Wisteria Floribunda Agglutinin

Check if the same lab product or an alternative is used in the 5 most similar protocols

The conjugation of Atto590 and Alexa594 with WFA was achieved through amine-reactive chemistry using the free amine groups in the WFA lectin. Freshly prepared 0.4 mg of Atto590 NHS ester (79636 and 68616, dye and protein labeling kit, respectively, Sigma Aldrich) or a full vial of Alexa Fluor™ 594 NHS ester (A10239, protein labeling kit, Thermo Fisher Scientific) dyes in DMSO was mixed with 3 mg or 1 mg, respectively, at a concentration not

< 2 mg / ml

(L8258, Sigma Aldrich) in bicarbonate buffer (pH 8.3). The reaction mixture was incubated in dark in a rotating mixer for 3 h at room temperature. The solution was then laid over on a silica column preactivated with phosphate buffered saline (PBS). The dye-conjugated WFA and the free dye were eluted with sodium phosphate buffer and were collected separately. Conjugation efficacy was tested by staining brain slices (1:150) and in vivo following intracranial injections (Fig. S1 in the Supplementary Material).

Benbenishty A., Peled-Hajaj S., Krishnaswamy V.R., Har-Gil H., Havusha-Laufer S., Ruggiero A., Slutsky I., Blinder P, & Sagi I. (2023). Longitudinal in vivo imaging of perineuronal nets. Neurophotonics, 10(1), 015008.

+ Open protocol

+ Expand

Cell Culture Reagents and Compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell culture reagents and chemicals were from Sigma Aldrich (St. Louis, MO, USA) or Thermo Fisher Scientific (Waltham, MA, USA) unless otherwise stated. Drugs and biologics as follows: ABT-737 (Selleck Chemicals); CHX (Sigma); YOYO3 and SYTO reagents (Life Technologies/Thermo Fisher Scientific), mTNFα and mTRAIL (Peprotech, Rocky Hill, NJ, USA); zVAD-fmk (ApexBio, Houston, TX, USA), birinapant (MedChem Express). Fluorophores: FITC (Cat. No. 46425, Thermo Fisher Scientific), Alexa Fluor 488 5-SDP Ester (Cat. No. A30052, Thermo Fisher Scientific), Alexa Fluor 594 NHS Ester (Cat. No. A37572, Thermo Fisher Scientific).

Gelles J.D., Mohammed J.N., Santos L.C., Legarda D., Ting A.T, & Chipuk J.E. (2019). Single-Cell and Population-Level Analyses Using Real-Time Kinetic Labeling Couples Proliferation and Cell Death Mechanisms. Developmental cell, 51(2), 277-291.e4.

+ 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?

Revolutionizing how scientists
search and build protocols!