Spectra por float a lyzer g2

Manufactured by Merck Group

Sourced in Germany, United States

The Spectra Por Float-A-Lyzer G2 is a dialysis device used for sample preparation and purification. It is designed to facilitate the dialysis process by providing a buoyant, easy-to-handle platform for membrane-based separation.

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Lab products found in correlation

Freezone plus 12, by Labconco (1 mentions) Cyanine7.5 nhs ester, by Lumiprobe (1 mentions)

Spelling variants of this product

Spectra/Por (8 citations) Float-A-Lyzer (6 citations) Spectra-Por Float-A-Lyzer (2 citations) Spectra-Por s Float-A-Lyzer s G2 (2 citations)

4 protocols using spectra por float a lyzer g2

Synthesis and Characterization of Carbon Nanodots

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Carbon nanodots were synthesized based on our published microwave-assisted method [9 (link),11 (link),12 (link)]. The samples were diluted with 10.0 mL of DDI water and then dialyzed against 5.0 L of DDI water using a 500–1000 Da MWCO (Spectra Por Float-A-Lyzer G2, 10 mL, G235063; MilliporeSigma; Darmstadt, Germany). The sample was dialyzed over three days under a gentle stir with a new DDI water replacement every 24 h. The resultant clear, brown-orange aqueous solution was lyophilized (Labconco FreeZone Plus 12; Kansas City, MO, USA) to produce the dried carbon nanodots. The Cary^® Eclipse™ Fluorescence Spectrophotometer was used for characterizing the photoluminescence of carbon nanodots.

Erikson K.M., El-Khouri K., Petric R., Tang C., Chen J., Vasquez D.E., Fordahl S.C, & Jia Z. (2023). Carbon Nanodots Attenuate Lipid Peroxidation in the LDL Receptor Knockout Mouse Brain. Antioxidants, 12(5), 1081.

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Production of Viral Vectors for Tracking and Transduction

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Viral vectors were produced by the Duke Viral Vector Core Facility (Department of Neurobiology, Duke University School of Medicine), as described previously (66 )(66 ). Briefly, HEK-293T cells grown in 10 cm plates were transfected using a calcium phosphate-based protocol with either 10 μg of psPAX2, 2.5 μg of pREV, 5 μg pMD2.G (VSV-G), and 5 μg of the eGFP.Vpr plasmid (described above) or 10 μg of psPAX2, 2.5 μg of pREV, 5 μg pMD2.G (VSV-G), 15 μg of the transgene/reporter gene pLenti-GFP, and 5 μg of the pmiRFP670.Vpr plasmid (described above), to produce VLPs suitable for single-particle tracking or transduction assays, respectively. In both cases, the media was replaced at 24 h after transfection, and 72 h post-transfection, supernatant containing pseudotyped particles was centrifuged at 450 × g for 10 min and filtered through a 0.45-μm pore-size filter.
Viral titers were determined using a p24 -enzyme-linked immunosorbent assay (ELISA) and reported in transducing units/ml (TU/mL), typically 2–4×10⁷ TU/mL. The virus solution was stored in aliquots at −80 °C. Prior to single-particle tracking and immunofluorescence experiments the virus solution was buffer exchanged by dialysis (Spectra-Por Float-A-Lyzer G2, MWCO 100 kDa, Millipore Sigma) against PBS (Genesee Scientific, Cat #: 25–507) at 4 °C.

Humphrey P.A., Wong A.J., Vogelstein B., Zalutsky M.R., Fuller G.N., Archer G.E., Friedman H.S., Kwatra M.M., Bigner S.H, & Bigner D.D. (1990). Anti-synthetic peptide antibody reacting at the fusion junction of deletion-mutant epidermal growth factor receptors in human glioblastoma.. Proceedings of the National Academy of Sciences of the United States of America, 87(11), 4207-4211.

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Protein Release from Nanohydrogels

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The release of loaded protein from the NanoC and NanoS was investigated in two different release media: plain PBS buffer (10 mM, pH 7.4, 136 mM NaCl) or PBS buffer supplemented with 10 mM GSH to simulate the extracellular and intracellular environment, respectively. The protein loaded nanohydrogel was suspended into release media at a concentration of 5 mg/mL and then the nanohydrogel suspension was transferred into a dialysis device with a Mw cutoff at 100 kDa (Spectra-Por^® Float-A-Lyzer^® G2, Sigma-Aldrich, St. Louis, MO, USA) in triplicate. The dialysis device was submerged into a falcon test tube with14 mL release medium, incubated at 37 °C and gently shaken at 50 rpm. At desired time intervals, 0.6 mL of release medium was sampled and replenished with an equal amount of fresh medium. The released protein was determined by Micro BCA assay. The released sample of GSH-containing medium was further purified by dialysis (Mw cutoff 3.5 kDa) against 0.01 M pH 7.4 PBS for 3 days before the Micro BCA assay to remove GSH that interferes with the colorimetric assay.

Deng S., Gigliobianco M.R., Mijit E., Minicucci M., Cortese M., Campisi B., Voinovich D., Battistelli M., Salucci S., Gobbi P., Lupidi G., Zambito G., Mezzanotte L., Censi R, & Di Martino P. (2021). Dually Cross-Linked Core-Shell Structure Nanohydrogel with Redox–Responsive Degradability for Intracellular Delivery. Pharmaceutics, 13(12), 2048.

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Synthesis of Cyanine7.5-labeled Phospholipid

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Cyanine7.5 NHS ester (Lumiprobe GmbH, Germany) was conjugated to the amine terminus of a commercially available phospholipid with a PEG2000 spacer (DSPE-PEG(2000)-NH₂) via a NHS-mediated coupling reaction to afford the target Cyanine7.5 dye-functionalised phospholipid (DSPE–PEG(2000)–Cyanine7.5) after purification by dialysis. DSPE–PEG(2000)–Cyanine7.5 (Fig. 1) was characterised by various analytical and spectroscopic techniques (see Supporting information, Figs. S1–S2). In a typical reaction, DSPE–PEG(2000)–NH₂ (3.0 mg, 1.1 μmol) and 6 μL of triethylamine were dissolved in 120 μL of dry DMSO. To this solution, 120 μL of DMSO containing Cyanine7.5 NHS ester (1.7 mg, 2.2 μmol) was added dropwise. The resulting mixture was allowed to react at room temperature overnight under continuous stirring. Distilled water was then added to the reaction mixture. The solution was centrifuged, and the supernatant was passed through a 0.45 μm filter to remove insoluble traces. The supernatant was then dialysed using a Spectra-Por^® Float-A-Lyzer^® G2 (Sigma-Aldrich, Milwaukee, Wis) (MW cutoff of 3.5–5 kDa) against water (3 × 500 mL). The dialysate, containing the pure product, was lyophilised and the residue dried in vacuo over P₂O₅. All lipids used in this study were purchased from Avanti Polar Lipids, Inc., USA.Fig. 1

Structure of the functionalised DSPE–PEG (2000)–Cyanine7.5 phospholipid.

Fig. 1

Lin S., Shah A., Hernández-Gil J., Stanziola A., Harriss B.I., Matsunaga T.O., Long N., Bamber J, & Tang M.X. (2017). Optically and acoustically triggerable sub-micron phase-change contrast agents for enhanced photoacoustic and ultrasound imaging. Photoacoustics, 6, 26-36.

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