F7250

Manufactured by Merck Group

Sourced in Germany

The F7250 is a laboratory equipment product manufactured by Merck Group. It serves as a general-purpose instrument used for various scientific applications. The core function of the F7250 is to perform measurements and analyses, providing accurate and reliable data for research and testing purposes.

Automatically generated - may contain errors

Lab products found in correlation

Plan apochromat 63x 1.40 oil dic m27 objective, by Zeiss (1 mentions) Polymorphprep, by Axis-Shield (1 mentions) Lsm 700 axio imager 2, by Zeiss (1 mentions) Cellsens dimension, by Olympus (1 mentions) Fluorescent conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Spinsr10 confocal system, by Olympus (1 mentions) Aqua poly mount, by Agilent Technologies (1 mentions) Dapi 4 6 diamidino 2 phenylindole, by Cell Signaling Technology (1 mentions) Fitc dextran, by Merck Group (1 mentions) Hanks balanced salt solution (hbss), by Welgene (1 mentions) Zeba spin desalting column, by Thermo Fisher Scientific (1 mentions) F4883, by Merck Group (1 mentions) Hitrap q ff, by GE Healthcare (1 mentions) Hitrap, by Cytiva (1 mentions) Tcs sp5 confocal microscope, by Leica (1 mentions) Hcx pl apo lambda blue 63x 1.4 oil objective, by Leica (1 mentions) Doxorubicin, by Merck Group (1 mentions) Bortezomib, by Merck Group (1 mentions) Pk pf647 1, by PromoCell (1 mentions) Carfilzomib, by Merck Group (1 mentions)

Close spelling variants of this product

F7250 Sigma (2 citations)

12 protocols using f7250

Measuring Phagocytosis of Aspergillus Conidia

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

Opsonization and phagocytosis was measured in an assay using FITC-conjugated conidia and isolated human neutrophils. FITC-conjugation was made by mixing FITC powder (F7250, Sigma-Aldrich) and conidia (5 × 10⁻⁸ μg/conidia) for 30 min end-over-end at room temperature followed by removal of unbound FITC by extensive washing. FITC-conjugated A. fumigatus conidia (1 × 10⁷/ml) were opsonized for 30 min at 37°C in 10% C1q-deficient serum including 10 μg/ml of either the MBL inhibitor (3F8) or mock-inhibitor (1C10). Opsonized conidia were washed and combined with human neutrophils isolated with Polymorphprep (Axis-Shield, Oslo, Norway) according to the manufacturer’s instructions. Neutrophils and conidia co-incubated for 30 min at 37°C in a cell ratio of 1:5. After washing the cells and before flow cytometric analysis, 50 μl tryphan blue was added to quench fluorescence from non-ingested conidia. FITC-positive neutrophils were identified by gating. Barbital buffer was used as dilution/washing buffer throughout the experiment.
We also performed fluorescence and differential interference contrast (DIC) imaging of neutrophils phagocytizing FITC-conjugated A. fumigatus conidia (using the same protocol) to get a visual impression of the process. We used Zeiss LSM 700 Axio Imager 2 with a Plan-Apochromat 63x/1.40 Oil DIC M27 objective and Carl Zeiss ZEN Blue edition software.

Rosbjerg A., Genster N., Pilely K., Skjoedt M.O., Stahl G.L, & Garred P. (2016). Complementary Roles of the Classical and Lectin Complement Pathways in the Defense against Aspergillus fumigatus. Frontiers in Immunology, 7, 473.

+ Open protocol

+ Expand

Immunofluorescence Imaging of GAS Infection

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

Cells infected with FITC (Sigma-Aldrich, F7250)-labelled GAS strains were washed with PBS, fixed for 15 min with 4% paraformaldehyde in PBS, permeabilized for 5 min in 0.2% Triton X-100 in PBS and blocked using 5% BSA for 1 h at room temperature. Then, cells were stained with indicated primary antibodies at 4 °C for overnight, followed by incubation with fluorescent-conjugated secondary antibodies (Jackson ImmunoResearch). Nuclei were counterstained with DAPI (4,6-diamidino-2-phenylindole) (Cell Signaling Technology, #4083). Slides were mounted using Aqua-Poly/Mount (Dako). Images were captured using a laser scanning confocal microscope (Olympus SpinSR10 Confocal System) with OLYMPUS cellSens Dimension. Z-stack images (optical slice 0.5 μm, 12 slices per 5.5-μm stack) were captured using a Zeiss 880 laser scanning confocal microscope (Zeiss LSM880) at 63× magnification and analyzed using Zeiss Zen software Blue edition.

Deng W., Bai Y., Deng F., Pan Y., Mei S., Zheng Z., Min R., Wu Z., Li W., Miao R., Zhang Z., Kupper T.S., Lieberman J, & Liu X. (2022). Streptococcal pyrogenic exotoxin B cleaves GSDMA and triggers pyroptosis. Nature, 602(7897), 496-502.

+ Open protocol

+ Expand

Imaging Fluorescent Beads and Dye Characterization

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

To characterize the general operation of our devices, we imaged fluorescent beads and fluorescein isothiocyanate (FITC) dye. Fluorescent beads: we employed Fluoresbrite 1.0 μm blue-green fluorescent microspheres (Polysciences catalog #17154) and FluoSpheres 0.5 μm red fluorescent microspheres (Invitrogen catalog # F8812). We diluted bead stock solutions 1:1 v/v in 1% sodium dodecyl sulfate to reduce clumping and then diluted this mixture 1:100 v/v in ethanol to allow rapid evaporation. We spread 10 µL of the ethanol mixture onto a 15 mm × 15 mm area of a 22 mm coverslip. Beads remain fixed after evaporation. FITC dye: we dissolved 20 mg of FITC (Sigma-Aldrich F7250) in 1 mL of ethanol and mixed 50 μL of this solution in 950 μL of water. We spread 2 μL of this mixture between a slide and a 22 mm coverslip.

Cai H., Wang Y.L., Wainner R.T., Iftimia N.V., Gabel C.V, & Chung S.H. (2019). Wedge prism approach for simultaneous multichannel microscopy. Scientific Reports, 9, 17795.

+ Open protocol

+ Expand

Intestinal Permeability Assay in Mice

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

We food and water-starved both control and treated mice of both strains overnight and kept them in a cage without bedding to limit the coprophagic behavior [16 (link)]. We measured mice body weights the following morning. We administered PBS dissolved FITC-dextran (100mg/ml, Cat#F7250, Sigma-Aldrich, Missouri, US) in each mouse (44mg/100g body weight) by oral gavaging following 4h of incubation. We anesthetized mice by isoflurane inhalation and collected blood using a 1ml syringe with a 25G needle by cardiac puncture. We isolated serum from the blood using the protocol described previously and diluted it with an equal volume of PBS. We added 100μl diluted serum in each well of a 96 well microplate in duplicate. We measured fluorescence at 528 nm (emission wavelength) by exciting at 485 nm (20 nm bandwidth at both excitation and emission) [17 ].

Mukhopadhyay S., Saha S., Chakraborty S., Prasad P., Ghosh A, & Aich P. (2022). Differential colitis susceptibility of Th1- and Th2-biased mice: A multi-omics approach. PLoS ONE, 17(3), e0264400.

+ Open protocol

+ Expand

FITC-Labeled Fibrinogen Purification

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

Fibrinogen from human plasma (Sigma-Aldrich, F-4883) was labeled with FITC (Sigma-Aldrich, F7250) according to the manufacturer’s recommendation. In brief, 20 mg of fibrinogen was incubated with 20-fold molar excess of FITC for 2 h at 4 °C with rotation. To remove free FITC, 20 mM Tris-Cl was added to the reaction and the mixture was desalted in DPBS [phosphate buffered saline (PBS) supplemented with 1 mM CaCl₂ and 1 mM MgCl₂; Hyclone] using Zeba™ Spin Desalting Columns (Thermo Fisher Scientific, 89892). Further purification was accomplished by using anion exchange chromatography column (HiTrap Q FF; GE Healthcare, 17–5053-01) to remove heavily labeled non-functional fibrinogen. The resulting fraction containing functional FITC-fibrinogen was then diluted with Hank’s Balanced Salt Solution (HBSS; WELGENE) containing 1.3 mM CaCl₂ and 1 mM MgSO₅. Due to the aggregation-prone nature of the labeled fibrinogen, labeling was performed immediately prior to each experiment.

Kim J., Lee J., Jang J., Ye F., Hong S.J., Petrich B.G., Ulmer T.S, & Kim C. (2020). Topological adaptation of transmembrane domains to the force-modulated lipid bilayer is a basis of sensing mechanical force. Current biology : CB, 30(9), 1614-1625.e5.

+ Open protocol

+ Expand

Labeling and Imaging Photoreceptor Outer Segments

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

Photoreceptor OS were obtained from InVision BioResources (WA, USA) or prepared in-house according to Molday et al., 198739 (link). OS labeling: Two mg/ml stock solution of FITC isomer 1 (F7250, Sigma) in 0,1M sodium bicarbonate at pH 9.0–9.5, was prepared, filter-sterilized, and stored in aliquots at −20 °C. For labeling with FITC, OS were suspended in solution containing 10% sucrose, 20 mM sodium phosphate, and 5 mM taurine, and incubated with FITC for 1,5 h at room temperature, rotating in the dark. FITC-labeled OS were washed 5 times in buffer containing 10% sucrose, 20 mM sodium phosphate and 5 mM taurine, suspended in DMEM, aliquoted and stored at −80 °C until use.
iPSC-RPE cells were incubated with OS for 2 h/37 °C, after which the OS were removed by triple washing with PBS. Cells were fixed by 4% PFA for 15 min/RT, washed and permeabilized by 0.1% Triton-X. Phallodin staining was performed for 30 min, the samples were then mounted with vectashield mounting media with DAPI. The samples were visualized by Leica confocal microscope TCS SP5 using HCX PL APO lambda blue 63X/ 1.4 OIL objective.

Lukovic D., Artero Castro A., Delgado A.B., Bernal M.D., Luna Pelaez N., Díez Lloret A., Perez Espejo R., Kamenarova K., Fernández Sánchez L., Cuenca N., Cortón M., Avila Fernandez A., Sorkio A., Skottman H., Ayuso C., Erceg S, & Bhattacharya S.S. (2015). Human iPSC derived disease model of MERTK-associated retinitis pigmentosa. Scientific Reports, 5, 12910.

+ Open protocol

+ Expand

Synthesis and Purification of FITC-Labeled Alendronate

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

Fluorescein isothiocyanate isomer I (1.13 μmol, F7250, Sigma) was dissolved in DMSO and then mixed with 11.3 μmol of alendronate dissolved in bicarbonate buffer, pH 9.0. The volume was diluted to 1 ml with distilled water, and the solution was incubated for 2 h at room temperature under constant mixing. To precipitate alendronate, 39.6 μmol of CaCl₂ was added, and the mixture was centrifuged (14,000 g, 10 min). The precipitate was washed with 1 ml of distilled water five times. To redissolve alendronate, 39.6 μmol of EDTA was added to the precipitate. Then, 100 μl of PBS was added until all precipitate dissolved, and the solution was shaken with ultrasonication for 30 min. The final solution (referred to as Aln-FITC) was lyophilized, and an orange powder was obtained containing FITC-labeled alendronate and free alendronate, which was quantified and purified by liquid chromatography.

Li Y., Fu Y., Zhang H., Song J, & Yang S. (2020). FITC-Labeled Alendronate as an In Vivo Bone pH Sensor. BioMed Research International, 2020, 4012194.

+ Open protocol

+ Expand

Annexin V Fluorescent Labeling Protocol

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

Annexin V was prepared following a published protocol^{28 (link)}, and coupled to either fluoresceinisothiocyanate (Sigma; F7250) or to PromoFluor 647 using its N-hydroxysuccinimidyl ester (PromoCell, Heidelberg, Germany; PK-PF647-1). Drugs used in this study were obtained from Sigma-Aldrich (melphalan; M2011), Selleck Chemicals, München, Germany (bortezomib; S1013, carfilzomib; S2853, nutlin-3A; S8059), and Merck, Darmstadt, Germany (doxorubicin; 324380). A solution of Na-resazurin (Sigma; R7017) in PBS was used as reagent in viability assays (alamarBlue).

Munawar U., Roth M., Barrio S., Wajant H., Siegmund D., Bargou R.C., Kortüm K.M, & Stühmer T. (2019). Assessment of TP53 lesions for p53 system functionality and drug resistance in multiple myeloma using an isogenic cell line model. Scientific Reports, 9, 18062.

+ Open protocol

+ Expand

FITC-Labeling of M. bovis BCG for Phagocytosis

Cited 1 time

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

M. bovis BCG (Tokyo 172 strain, Japan BCG laboratory #876391) was labeled with FITC (SIGMA #F7250), as described previously^{99 (link)}. Briefly, 6.0 × 10⁷ CFU of M. bovis BCG were resolved in 200 μl of 0.1 M carbonate–bicarbonate buffer (pH 9.6)/0.05% Tween 80 containing of 30 μg/ml FITC, and incubated at 37 °C for 15 min. The FITC-labeled mycobacteria were washed with PBS/0.05% Tween 80. For phagocytosis assay, WT or Trem2^−/− BMDMs (1 × 10⁵) were incubated with 1 × 10⁶ (MOI 10:1) of FITC-labeled mycobacteria in a 96-well plate for 4 h at 37 °C. Negative controls were prepared in identical conditions but incubated on ice (0 °C). The phagocytosis was stopped by washing with ice-cold PBS. The phagocytosed mycobacteria were analyzed by flow cytometer after addition of 0.02% Trypan Blue to quench FITC fluorescence of extracellular but not internalized mycobacteria¹⁰⁰.

Iizasa E., Chuma Y., Uematsu T., Kubota M., Kawaguchi H., Umemura M., Toyonaga K., Kiyohara H., Yano I., Colonna M., Sugita M., Matsuzaki G., Yamasaki S., Yoshida H, & Hara H. (2021). TREM2 is a receptor for non-glycosylated mycolic acids of mycobacteria that limits anti-mycobacterial macrophage activation. Nature Communications, 12, 2299.

+ Open protocol

+ Expand

Protein and Starch Visualization in Noodles

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

The morphology of protein/starch matrices (wheat flours and the inner and outer section of cooked noodles) was observed under an LSM 700 confocal microscope (Carl Zeiss, Germany) as described previously (Bhattarai et al., 2016) . Fluorescamine (F9015, Sigma) and fluorescein isothiocyanate (FITC; F7250, Sigma) were used for labelling protein and starch, respectively.

Li H.T., Li Z., Fox G.P., Gidley M.J, & Dhital S. (2021). Protein-starch matrix plays a key role in enzymic digestion of high-amylose wheat noodle. Food chemistry, 336.

+ 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!