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Fluorescamine

Fluorescamine is a fluorogenic agent used to label and detect primary amines, including amino acids, peptides, and proteins.
It reacts with primary amines to produce a highly fluorescent compound, allowing for sensitive detection and quantification of these compounds.
Fluorescamine has been widely used in biochemical and analytical applications, such as amino acid analysis, protein determination, and cellular labeling.
The PubCompare.ai platform helps researchers optimize Fluorescamine protocols by locating and comparing methods from literature, pre-prints, and patents, identifying the best protocols and products to enhance reproducibility and research accuarcy.

Most cited protocols related to «Fluorescamine»

Amyloid Aggregation Kinetics Assay

Cited 6 times

For Aβ40, HFIP-treated powder was dissolved in 10 mM NaOH. Then eight volumes of PBS buffer and one volume of 10 mM HCl solution were added, in that order. Aβ40 sample was mixed with various concentrations of ThT and then transferred to microplate for kinetics studies. The final concentration of Aβ40 was 50 µM.
For Aβ42, HFIP-treated powder was dissolved in CG buffer, and was then buffer exchanged to PBS using a 5 ml HiTrap desalting column (GE healthcare). The concentration of Aβ42 was determined using fluorescamine [40 (link)], and hen egg white lysozyme was used to prepare the standard curve. Aβ42 was then diluted to 10 µM in PBS buffer containing various concentrations of ThT. In an alternative protocol, HFIP-treated Aβ42 was dissolved in CG buffer to 200 µM concentration, and was then diluted 20-fold to PBS buffer containing various concentrations of ThT. The final Aβ42 concentration was also 10 µM.
Ure2 protein was dissolved in PG buffer to 200 µM concentration, and was then diluted 20-fold to PBS buffer containing various concentrations of ThT. The final concentration of Ure2 was 10 µM.
The volume of aggregation was 50 µl for all the samples. Samples were kept on ice whenever possible. All samples were transferred to a black 384-well Nonbinding Surface microplate with clear bottom (Corning product# 3655), and sealed with a polyester-based sealing film (Corning product# PCR-SP). To begin aggregation, the microplate was transferred to a Victor 3 V plate reader (Perkin Elmer) and was incubated at 37°C without agitation. The ThT fluorescence was measured through the bottom of the plate approximately every 5 min, with excitation filter of 450 nm and emission filter of 490 nm.

Xue C., Lin T.Y., Chang D, & Guo Z. (2017). Thioflavin T as an amyloid dye: fibril quantification, optimal concentration and effect on aggregation. Royal Society Open Science, 4(1), 160696.

Publication 2017

Buffers Fluorescamine Fluorescence hen egg lysozyme Kinetics Polyesters Powder Proteins

Quantifying Amyloid-beta Peptide Concentration

Cited 5 times

One tube of Aβ40 powder (approx. 200 µg) was dissolved in 500 µl of HFIP, split into two tubes, and dried with a gentle stream of argon gas. For the absorbance method, one tube of Aβ40 was then dissolved in CG buffer (20 mM CAPS, 7 M GdnHCl, pH 11). The volume of CG buffer was chosen so that the absorbance reading would be between 0.5 and 1.0. The sample was used for absorbance measurement without further dilution. Absorbance at 280 nm was measured using a quartz microcuvette with 1 cm path length on a Jasco V-630 spectrophotometer. Concentration was calculated using an extinction coefficient of 1280 M⁻¹ cm⁻¹ [5 (link)]. The other tube of Aβ40 was dissolved in one volume of 10 mM NaOH. Then eight volumes of PBS were added, followed by one volume of 10 mM HCl for neutralization. To measure concentration using fluorescamine, 5 µl of Aβ40 sample was mixed with 40 µl of PBS and 5 µl of fluorescamine stock solutions. The sample was further diluted if needed so that fluorescence intensity readings would fall in the middle of the standard curves. For the fluorescamine method with DMSO or HFIP, 35 µl of DMSO or HFIP were used to substitute 35 µl of PBS buffer. Fluorescence measurements were performed the same way as the standard curves. Concentration was determined using the five different fluorescamine concentrations, with the standard curves that were established above.

Xue C., Lee Y.K., Tran J., Chang D, & Guo Z. (2017). A mix-and-click method to measure amyloid-β concentration with sub-micromolar sensitivity. Royal Society Open Science, 4(8), 170325.

Publication 2017

Argon Buffers Extinction, Psychological Fluorescamine Fluorescence Powder Quartz Sulfoxide, Dimethyl Technique, Dilution

Peptide Degradation Assay in Baf/BCR-ABL Cells

Cited 5 times

A Baf/BCR-ABL cell pellet was washed with and resuspended in phosphate buffered saline (PBS; 137 mM NaCl, 10 mM Na₂HPO₄, 27 mM KCl, 1.75 mM KH₂PO₄, pH 7.4). The cells were submerged in liquid nitrogen for 1 min and rapidly thawed at 37 °C for a total of three cycles. The mixture was centrifuged at 14,000 x g for 5 min at 4 °C. The supernatant was transferred to a clean centrifuge tube and maintained on ice until use in the assay. Total protein concentration in the supernatant was measured using fluorescamine.^{42 (link)} Briefly, fluorescamine (10 μL, 3 mg/mL in acetone) was added to a cell lysate (30 μL) and incubated for 5 min at 25 °C. Fluorescence was measured with a fluorescence plate reader (SpectraMax M5, Molecular Devices, Sunnyvale, CA) with an excitation of 390 nm (bandwidth of 9 nm) and emission of 475 nm (bandwidth of 15 nm).
Assay of peptide degradation was performed by mixing peptide (1 μM) with the Baf/BCR-ABL cell lysate (3 mg/mL total cell protein) and incubating at 37 °C. Aliquots were removed from the reaction mixture at various time intervals. The reactions were stopped by adding HCl to a final concentration of 100 mM. A 0 min timepoint was prepared by adding the HCl to the lysate prior to addition of the substrate peptide. Reaction mixtures were then separated by capillary electrophoresis and detected by LIF. Peptide fragments were identified by adding standards (250 nM) to the HCl-terminated aliquots and comparing the electropherograms with and without the added standard. The average initial degradation and fragmentation rates were calculated using the first two time points by monitoring the change in peptide amount divided by the change in time per amount cytosolic protein. The units are defined as zmol of peptide per pg of cytosolic protein per s, or zmol pg⁻¹ s⁻¹.

Proctor A., Wang Q., Lawrence D.S, & Allbritton N.L. (2012). Metabolism of Peptide Reporters in Cell Lysates and Single Cells. The Analyst, 137(13), 3028-3038.

Publication 2012

Acetone Biological Assay Cells Cytosol Electrophoresis, Capillary Fluorescamine Fluorescence Medical Devices Nitrogen Peptide Fragments Peptides Phosphates Proteins Saline Solution Sodium Chloride

Characterization and Encapsulation of PAMPs in Ace-DEX Microparticles

Cited 4 times

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Publication 2019

Acetals Anabolism aquasil Biological Assay Chlorides cyclic guanosine monophosphate-adenosine monophosphate Deuterium Deuterium Oxide Dextran 70 Ethanol Fluorescamine High-Performance Liquid Chromatographies Imiquimod Medical Devices Methanol murabutide Pathogen-Associated Molecular Pattern Molecules Poly I-C Solvents Spectroscopy, Nuclear Magnetic Resonance Sulfoxide, Dimethyl

Multiparametric Cell Cytotoxicity Assay

Cited 4 times

Following the treatment, cells were washed three times with calcium and magnesium-containing PBS to remove the debris. Cells were solubilized using 200 µL 0.1% Triton X-100 non-ionic detergent containing borate buffer (pH 9.2) supplemented with 10 mM EDTA and were placed on a shaker for 5 min. The microplate cytotoxicity assay is based on a multiparametric measurement [10 (link)]. ATP was determined by the luciferin/luciferase technique [10 (link)] adapted for microplate method. To measure the ATP content, lysed samples were transferred into 96-well white optiplates, 20 µL/well each. 200 µL of dissolved ATP reagent was added to the wells. The working concentration of PI staining was 2 µg/mL in PBS. The extracted samples were incubated for 5 min at room temperature in the dark, and after a short shaking they were measured on plate reader (Enspire Multimode reader, Perkin Elmer, (Waltham, Massachusetts) at λexc 530 nm and λem 620 nm wavelengths. Fluorescence intensity of GFP was measured at λexc 480 nm and λem 520 nm wavelengths. We used fluorescamine (0.3 mg/mL dissolved in acetone) to determine the intracellular protein content. Bovine serum albumin standard was used to generate linear regression calibration curve. Protein concentration was determined in the range of 20–100 mg/L. 20 µL/well borate buffer-lysed sample/standard was transferred into a 96-well standard plate. After pipetting 150 µL of 0.1% Triton X-100 containing borate buffer into the wells, 50 µL of fluram/acetone was rapidly added to each well. After a short shaking, fluorescence intensity was measured on the plate reader at λexc 385 nm and λem 490 nm wavelengths.

Csepregi R., Temesfői V., Poór M., Faust Z, & Kőszegi T. (2018). Green Fluorescent Protein-Based Viability Assay in a Multiparametric Configuration. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(7), 1575.

Publication 2018

Acetone Biological Assay Borates Buffers Calcium Cells Cytotoxin Detergents Edetic Acid Fluorescamine Fluorescence Fluram Luciferases Luciferins Magnesium Proteins Protoplasm Serum Albumin, Bovine Triton X-100

Most recents protocols related to «Fluorescamine»

Comprehensive Analysis of HECP2k Polymer

The synthesis of HECP2k was confirmed by ¹H NMR and ¹³C NMR, respectively (600 MHz, AVANCE 600, Brucker, Germany). D₂O was used as the solvent. Each sample was prepared at a concentration of 10 mg/mL for ¹H NMR and 50 mg/mL for ¹³C NMR.
The synthesis of the polymers was also confirmed using an ATR (attenuated total reflectance) FT-IR spectrometer (Nicolet 6700, Thermo Scientific, Waltham, MA, USA). The measurements were performed in the wavenumber range of 4000–650 cm⁻¹ with 8 cm⁻¹ by 32 times.
The primary amine (1° amine) quantification of HECP2k was performed via fluorescamine assay. The calibration curve was prepared by using PEI2k as a standard. The primary amine contents of the PEI2k (molar amounts of 1°:2°:3° amine = 40:36:24) were provided by the manufacturer. We reacted 75 μL of HECP2k sample solution (10 μg/mL, PBS) and 25 μL of fluorescamine solution (3 mg/mL, DMSO) for 15 min at 25 °C. The fluorescence was measured at Ex/Em = 380/470 nm with a microplate reader (Synergy H1, BioTek, Winooski, VT, USA). After the measurements, the molar amount of primary amine per 1 g of HECP2k was calculated for each sample and compared with ¹H NMR results.
The molecular weights of the polymers were measured via gel permeation chromatography (GPC, YL-9100 HPLC System, Youngin Chromass, Anyang, Korea). Each sample was prepared at a concentration of 10 mg/mL. We used 1% formic acid as an eluent. Poly(ethylene glycol)s with various molecular weights were used as standards for analysis. The assay was run on an Ultrahydrogel 250 column (Waters, Milford, MA, USA) at 0.6 mL/min of flow rate.

Park J., Kim S, & Kim T.I. (2023). Polyethylenimine-Conjugated Hydroxyethyl Cellulose for Doxorubicin/Bcl-2 siRNA Co-Delivery Systems. Pharmaceutics, 15(2), 708.

Publication 2023

1H NMR Amines Anabolism Biological Assay Carbon-13 Magnetic Resonance Spectroscopy Fluorescamine Fluorescence formic acid Gel Chromatography High-Performance Liquid Chromatographies Molar PER1 protein, human Polyethylene Glycols Polymers Solvents Sulfoxide, Dimethyl

Characterizing Macroporous Hydrogel Structure

The structure of the swollen macroporous hydrogel samples was evaluated by confocal laser scanning microscopy using a Nikon TE-2000 inverted microscope equipped with an EZ-C1 confocal laser (Nikon, Tokyo, Japan). For this purpose, the macroporous hydrogel samples were swollen in PBS (pH 7.4) and stained with fluorescamine (0.3 μg/mL in acetone) to provide amino-specific staining. The excitation wavelength was 408 nm and fluorescence signals were collected at 515±30 nm. Image analysis software (ImageJ, National Institutes of Health, USA) was used for 3D reconstruction of the macroporous hydrogel structure and to determine the mean pore size. An effective pore diameter (d) was calculated as follows:
where L is a pore long axis length and S is a pore short axis length. The mean pore size was determined by randomly measuring at least 100 pores for each macroporous hydrogel sample.

Tolstova T., Drozdova M., Popyrina T., Matveeva D., Demina T., Akopova T., Andreeva E, & Markvicheva E. (2023). Preparation and In Vitro Evaluation of Chitosan-g-Oligolactide Based Films and Macroporous Hydrogels for Tissue Engineering. Polymers, 15(4), 907.

Publication 2023

Acetone Epistropheus Fluorescamine Fluorescence Hydrogels Microscopy Microscopy, Confocal Reconstructive Surgical Procedures

Fluorescamine-based pH-MUC1 NP Assay

The pH-MUC1 NP solution was diluted with three solutions of PBS buffer at pH 7.4, 6.8, and 5.5, respectively, to achieve a final concentration of 1.6 mg pH-MUC1 NP/mL. Next, 10 μL of fluorescamine solution in DMSO (3 mg/mL) was mixed with 150 μL of each NP solution. Each solution was then transferred into a black flat bottom 96-well plate and analyzed using a plate reader (Spectra Max M3 by Molecular devices). PBS solutions at pH 7.4, 6.8, and 5.5 and PBS at pH 7.4, 6.8, and 5.5 with pH-MUC1 NPs without fluorescamine were used as controls.

Wlodarczyk M.T., Dragulska S.A., Chen Y., Poursharifi M., Acosta Santiago M., Martignetti J.A, & Mieszawska A.J. (2023). Pt(II)-PLGA Hybrid in a pH-Responsive Nanoparticle System Targeting Ovarian Cancer. Pharmaceutics, 15(2), 607.

Publication 2023

Buffers Fluorescamine Medical Devices MUC1 protein, human Sulfoxide, Dimethyl

Double-Labeling Method for Carbohydrate and Protein Imaging

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Publication 2023

acetonitrile Argon Ion Lasers Borates Buffers Carbohydrates Centrifugation Fluorescamine Fluorescence Glycerin Krypton Microscopy Microscopy, Confocal Nails Periodic Acid Periodic Acid-Schiff Reaction Potassium Proteins Schiff's reagent Sodium Sulfites Tetragonopterus

Functionalization of GO-POSS with Glutathione

Maleic anhydride was first attached onto the GO–POSS surface via an amidation reaction [49 (link)]. Then, 1.0 g of GO–POSS was dispersed into 50 mL of toluene solution, and maleic anhydride was added into the system and refluxed at 160 °C under an N₂ atmosphere for 2 h. Next, the toluene solvent was removed via rotary evaporation, and the GO–POSS nanoparticles modified with maleic anhydride were washed with deionized water and dried in an oven at 60 °C for 24 h.
Glutathione (GSH) was then used to modify the GO–POSS via a Michael addition reaction to obtain the final GPG composite product. Next, 100 mg of maleic-anhydride-modified GO–POSS nanosheets was dispersed in a PBS solution (0.1 mol L⁻¹, pH 6.0), 100 mg of glutathione was added into the suspension, and the resulting mixture was incubated in PBS solution for 48 h. After centrifugation to remove the supernatant, the product was washed with deionized water and dried under a vacuum at 60 °C for 12 h.
The GSH content on the GPG composite was determined via the fluorescamine method [50 (link),51 (link)] by recording the fluorescence intensity at 478 nm with excitation at 392 nm.

Sun J., Jia L, & Chen X. (2023). Efficient Adsorption and Extraction of Glutathione S-Transferases with Glutathione-Functionalized Graphene Oxide–Polyhedral Oligomeric Silsesquioxane Composite. Molecules, 28(1), 340.

Publication 2023

Atmosphere Centrifugation Fluorescamine Fluorescence Maleic Anhydride Solvents Toluene Vacuum

Top products related to «Fluorescamine»

Fluorescamine by Merck Group

Sourced in United States, Germany, United Kingdom, Switzerland, Israel, China

Fluorescamine is a chemical reagent used in analytical chemistry and biochemistry. It reacts with primary amines, such as those found in amino acids and proteins, to produce a fluorescent product that can be detected and quantified using fluorescence spectroscopy.

Bovine serum albumin (bsa) by Merck Group

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Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.

Dimethyl sulfoxide (dmso) by Merck Group

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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.

Fetal bovine serum (fbs) by Thermo Fisher Scientific

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

Enspire multimode plate reader by PerkinElmer

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The EnSpire Multimode Plate Reader is a versatile laboratory instrument designed for high-performance detection and analysis of a wide range of assays. It features multiple detection modes, including absorbance, fluorescence, and luminescence, enabling researchers to conduct a variety of experiments on a single platform.

Fluorescamine by Thermo Fisher Scientific

Sourced in United States

Fluorescamine is a fluorogenic reagent used for the detection and quantification of primary amines, including amino acids, peptides, and proteins. It reacts with primary amines to produce a highly fluorescent product, enabling sensitive and specific detection of these compounds.

Triton x 100 by Merck Group

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Triton X-100 is a non-ionic surfactant commonly used in various laboratory applications. It functions as a detergent and solubilizing agent, facilitating the solubilization and extraction of proteins and other biomolecules from biological samples.

Micro bca protein assay kit by Thermo Fisher Scientific

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The Micro BCA Protein Assay Kit is a colorimetric detection method for the quantification of protein concentrations. The kit utilizes the bicinchoninic acid (BCA) reaction to determine the total protein content of a sample.

Dulbecco s modified eagle s medium dmem by Thermo Fisher Scientific

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Dulbecco's modified Eagle's medium (DMEM) is a cell culture medium commonly used for the in vitro cultivation of various cell types. It provides a balanced salt solution, amino acids, vitamins, and other nutrients required for cell growth and maintenance.

Penicillin streptomycin by Thermo Fisher Scientific

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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.

More about "Fluorescamine"

Fluorescamine is a versatile fluorogenic agent that has been widely used in biochemical and analytical applications.
This highly reactive compound can label and detect primary amines, including amino acids, peptides, and proteins, by producing a highly fluorescent compound upon reaction.
This allows for the sensitive detection and quantification of these important biomolecules.
Fluorescamine has been employed in a variety of applications, such as amino acid analysis, protein determination, and cellular labeling.
The PubCompare.ai platform helps researchers optimize Fluorescamine protocols by locating and comparing methods from the literature, preprints, and patents, identifying the best protocols and products to enhance reproducibility and research accuracy.
When using Fluorescamine, researchers may also utilize other common laboratory reagents and techniques.
For example, Bovine Serum Albumin (BSA) is often used as a protein standard, while DMSO (Dimethyl Sulfoxide) can be used as a solvent for Fluorescamine stock solutions.
Fetal Bovine Serum (FBS) and Dulbecco's Modified Eagle's Medium (DMEM) are commonly used in cell culture applications, and Triton X-100 can be used as a detergent to solubilize proteins.
Specialized equipment, such as the EnSpire Multimode Plate Reader, may be employed to measure the fluorescence generated by the Fluorescamine-labeled compounds.
Additionally, the Micro BCA Protein Assay Kit can be used in conjunction with Fluorescamine to determine protein concentrations.
By incorporating these related terms and techniques, researchers can further optimize their Fluorescamine-based protocols and enhance the reproducibility and accuracy of their research.
The PubCompare.ai platform provides a valuable resource for identifying the best methods and products to streamline this process.