Polyethylene Glycol 300 is a low molecular weight, water-soluble polymer with a versatile range of applications in biomedical research and pharmaceutical formulations.
This nontoxic, odorless, and colorless compound is commonly used as a solvent, carrier, and excipient, owing to its excellent solubilizing and stabilizing properties.
Polyethylene Glycol 300 is particularly useful in the development of drug delivery systems, tissue engineering, and as a cryoprotectant for biomolecules.
Researchers can leverage the power of PubCompare.ai's AI-driven platform to optimize their Polyethylene Glycol 300 studies, locating the best protocols from literature, preprints, and patents, while experiencing data-driven decisions and enhanced reproducibility and accuracy.
Most cited protocols related to «Polyethylene glycol 300»
CUT&RUN begins with crude nuclei prepared according to published procedures. The following protocol is provided in step-by-step format (Appendix 2). Nuclei from ~5 × 108 cells at OD600 ~0.7 were prepared as described (Orsi et al., 2015 (link)), divided into 10 600 µL aliquots, snap-frozen and held at −80°C, then thawed on ice before use. Bio-Mag Plus Concanavalin A (lectin) coated beads were equilibrated with HNT (20 mM HEPES pH7.5, 100 mM NaCl and 0.1% Tween 20) that was supplemented with 1 mM each MgCl2, CaCl2 and MnCl2. Only Ca++ and Mn++ are needed to activate lectins, and omitting MgCl2 had no effect on binding of permeabilized cells to beads. The beads (300 µL) were rapidly mixed with a thawed nuclei aliquot and held at room temperature (RT) ≥5 min, placed on a magnet stand to clear (<1 min), and decanted on a magnet stand. The beads were then incubated 5 min RT in HNT supplemented with protease inhibitors (Roche Complete tablets) and 1 mM phenylmethylsulfonyl fluoride (PMSF) (=HNT-PPi) containing 3% bovine serum albumen (BSA) and 2 mM EDTA pH 8, then incubated 5’ with HNT-PPi +0.1% BSA (blocking buffer), using the magnet stand to decant. The beads were incubated 2 hr at 4°C with mouse anti-FLAG antibody (1:200–1:350), decanted, washed once in HNT + PMSF, then incubated 1 hr at 4°C with rabbit anti-mouse IgG antibody (1:200) in blocking buffer. The beads were washed once in HNT + PMSF, then incubated 1 hr at 4°C with pA-MN (600 µg/ml, 1:200) in blocking buffer. The beads were washed twice in HNT + PMSF and once in 20 mM HEPES pH 7.5, 100 mM NaCl (Digestion buffer), optionally including 10% polyethylene glycol 8000 for Sth1 and Mot1. The beads were brought up in 1.2 ml Digestion buffer, divided into 8 × 150 µL aliquots, equilibrated to 0°C, then quickly mixed with CaCl2, stopping the reaction with 150 µL 2XSTOP [200 mM NaCl, 20 mM EDTA, 4 mM EGTA, 50 µg/ml RNase A (Thermo Scientific, Waltham, MA, Catalog #EN0531) and 40 µg/ml glycogen (Sigma, Catalog #10901393001), containing 5-50 pg/ml heterologous mostly mononucleosome-sized DNA fragments extracted from formaldehyde crosslinked MNase-treated Drosophila chromatin as a spike-in]. After incubating at 37°C for 20 min, the beads were centrifuged 5 min at 13,000 rpm at 4°C, the supernatant was removed on a magnet stand and mixed with 3 µL 10% SDS and 2 µL Proteinase K (Invitrogen, Carlsbad, CA, Catalog #25530049), incubated at 70°C 10 min, then extracted at room temperature once with buffered phenol-chloroform-isoamyl alcohol (25:24:1, Sigma P2069), transferred to a phase-lock tube (Qiagen, Hilden, Germany, Catalog #129046), re-extracted with one vol CHCl3, transferred to a fresh tube containing 2 µL 2 mg/ml glycogen, precipitated by addition of 2–2.5 vol ethanol, chilled in ice and centrifuged 10 min at 13,000 rpm at 4°C. The pellet was rinsed with 100% ethanol, air-dried and dissolved in 25 µL 0.1 x TE8 (=1 mM Tris pH 8, 0.1 mM EDTA). To extend CUT&RUN for high-salt extraction, digestions were performed in a 50 µL volume, stopped with 50 µL 2XSTOP, omitting RNase and substituting the standard 200 mM NaCl with 4 M NaCl. After 20 min at 37°C, 200 µL 67 µg/ml RNase A was added, incubated 20 min, then centrifuged 13,000 rpm to clarify the supernatant.
Skene P.J, & Henikoff S. (2017). An efficient targeted nuclease strategy for high-resolution mapping of DNA binding sites. eLife, 6, e21856.
Full details of experimental procedures are provided in the supplementary materials. SpyCas9 and its point mutants were expressed in Escherichia coli Rosetta 2 strain and purified essentially as described (8 (link)). SpyCas9 crystals were grown using the hanging drop vapor diffusion method from 0.1 M tris-Cl (pH 8.5), 0.2 to 0.3 M Li2SO4, and 14 to 15% (w/v) PEG 3350 (polyethylene glycol, molecular weight 3350) at 20°C. Diffraction data were measured at beamlines 8.2.1 and 8.2.2 of the Advanced Light Source (Lawrence Berkeley National Laboratory), and at beamlines PXI and PXIII of the Swiss Light Source (Paul Scherrer Institut) and processed using XDS (50 (link)). Phasing was performed with crystals of selenomethionine (SeMet)–substituted SpyCas9 and native Cas9 crystals soaked individually with 10 mM Na2WO4, 10 mM CoCl2, 1 mM thimerosal, and 1 mM Er(III) acetate. Phases were calculated using autoSHARP (51 (link)) and improved by density modification using Resolve (52 (link)). The atomic model was built in Coot (53 (link)) and refined using phenix.refine (54 (link)). A. naeslundii Cas9 (AnaCas9) was expressed in E. coli Rosetta 2 (DE3) as a fusion protein containing an N-terminal His10 tag followed by MBP and a TEV (tobacco etch virus) protease cleavage site. The protein was purified by Ni-NTA (nickel–nitrilotriacetic acid) and heparin affinity chromatography, followed by a gel filtration step. Crystals of native and SeMet-substituted AnaCas9 were grown from 10% (w/v) PEG 8000, 0.25 M calcium acetate, 50 mM magnesium acetate, and 5 mM spermidine. Native and SeMet single-wavelength anomalous diffraction (SAD) data sets were collected at beamline 8.3.1 of the Advanced Light Source, processed using Mosflm (55 (link)), and scaled in Scala (56 (link)). Phases were calculated in Solve/Resolve (52 (link)), and the atomic model was built in Coot and refined in Refmac (57 (link)) and phenix.refine (54 (link)). For biochemical assays, crRNAs were synthesized by Integrated DNA Technologies, and tracrRNA was prepared by in vitro transcription as described (8 (link)). The sequences of RNA and DNA reagents used in this study are listed in table S2. Cleavage reactions were performed at room temperature in reaction buffer [20 mM tris-Cl (pH 7.5), 100 mM KCl, 5 mM MgCl2, 5% glycerol, 1 mM dithiothreitol] using 1 nM radio-labeled dsDNA substrates and 1 nM or 10 nM Cas9:crRNA:tracrRNA. Cleavage products were resolved by 10% denaturing (7 M urea) PAGE and visualized by phosphorimaging. Cross-linked peptide-DNA heteroconjugates were obtained by incubating 200 pmol of catalytically inactive (D10A/H840A) Cas9 with crRNA:tracrRNA guide and 10-fold molar excess of BrdU containing dsDNA substrate for 30 min at room temperature, followed by irradiation with UV light (308 nm) for 30 min. S1 nuclease and phosphatase–treated tryptic digests were analyzed using a Dionex UltiMate3000 RSLCnano liquid chromatograph connected in-line with an LTQ Orbitrap XL mass spectrometer equipped with a nanoelectrospray ionization source (Thermo Fisher Scientific). For negative-stain EM, apo-SpyCas9, SpyCas9: RNA, and SpyCas9:RNA:DNA complexes were reconstituted in reaction buffer, diluted to a concentration of ~25 to 60 nM, applied to glow-discharged 400-mesh continuous carbon grids, and stained with 2% (w/v) uranyl acetate solution. Data were acquired using a Tecnai F20 Twin transmission electron microscope operated at 120 keV at a nominal magnification of either ×80,000 (1.45 Å at the specimen level) or ×100,000 (1.08 Å at the specimen level) using low-dose exposures (~20 e− Å−2) with a randomly set defocus ranging from −0.5 to −1.3 μm. A total of 300 to 400 images of each Cas9 sample were automatically recorded on a Gatan 4k × 4k CCD (charge-coupled device) camera using the MSI-Raster application within the automated macromolecular microscopy software Leginon (58 (link)). Particles were preprocessed in Appion (45 (link)) before projection matching refinement with libraries from EMAN2 and SPARX (59 (link), 60 (link)) using RCT reconstructions (34 (link)) as initial models.
Jinek M., Jiang F., Taylor D.W., Sternberg S.H., Kaya E., Ma E., Anders C., Hauer M., Zhou K., Lin S., Kaplan M., Iavarone A.T., Charpentier E., Nogales E, & Doudna J.A. (2014). Structures of Cas9 Endonucleases Reveal RNA-Mediated Conformational Activation. Science (New York, N.Y.), 343(6176), 1247997.
Protein Expression and Purification—The pRSETa vector containing GCaMP2 was a kind gift of Karel Svoboda (Janelia Farm Research Campus, Howard Hughes Medical Institute). GCaMP2 was expressed and purified as described previously (17 ). Briefly, BL21(DE3) cells containing pRSETa harboring gcamp2 or gcamp2 mutants were grown in ZYM-5052 medium (18 ) for 48 h at 25 °C with shaking at 200 rpm. After centrifugation, cell lysis, and clarification, proteins were purified from the cell-free extract by nickel-affinity chromatography. Protein purity over 95% was confirmed by SDS-PAGE analysis. Proteins were dialyzed into 20 mm Tris-HCl, 100 mm NaCl, 2 mm CaCl2, pH 8.0, and concentrated. Calcium-free samples were prepared identically, except the buffer contained 5 mm EGTA instead of CaCl2. Crystallization and Data Collection—All GCaMP crystallization was carried out at 20 °C. All GCaMP protein samples for crystallization were in 20 mm Tris, 100 mm NaCl, 2 mm CaCl2, pH 8.0, except for the 8EF-apo mutant where the same buffer with 5 mm EGTA substituted for CaCl2 was used. All crystals used for data collection were grown using the hanging-drop vapor diffusion method in 24-well VDX plates. Crystallization of Ca2+-saturated dimeric GCaMP2 was described previously (17 ). The calcium-saturated K378W (at 5.6 mg/ml) and G87R (at 1.5 mg/ml) mutants crystallized ∼4 days after mixing with a precipitant solution consisting of 0.1 m magnesium formate dihydrate and 15% polyethylene glycol 3,350 using drop ratios of 2 μl of protein to 2 μl of precipitant for K378W and 1.5-2.5 μl for G87R. Ca2+-saturated monomeric GCaMP2 was crystallized identically to the K378W and G87R mutants using a drop ratio of 2 μl to 2 μl except that the drops were microseeded by streak seeding from K378W crystals immediately following setup. These crystals required more than 4 weeks to grow and had a distinct morphology. 8EF-apo GCaMP2 was crystallized after 1 week by mixing 2 μl of protein solution (9.5 mg/ml) with 2 μl of a precipitant solution consisting of 0.2 m lithium sulfate monohydrate, 0.1 m BisTris, pH 5.5, and 25% polyethylene glycol 3,350. All crystals were cryoprotected for data collection by quickly (<10 s) soaking in the precipitant solution supplemented with 20% glycerol and then mounted in a nitrogen gas stream at 100 K or plunged into liquid nitrogen for storage and transport to synchrotron beamlines. All data were collected at 100 K in a N2 gas stream. X-ray diffraction data for the G87R mutant was collected in-house on a Rigaku RU-H3R rotating copper anode x-ray generator, equipped with a Saturn 92 CCD detector and X-stream 2000 low-temperature system. Data for Ca2+-saturated monomeric GCaMP2 was collected at the Advanced Light Source, beamline 8.2.2. Diffraction data from crystals of Ca2+-dimer, K378W, and 8EF-apo GCaMP2 were collected at the Advance Photon Source, beamline 31-ID. X-ray diffraction data for G87R were integrated and scaled using d*TREK (19 (link)) from within the CrystalClear software package (Rigaku/Molecular Structure Corporation, Woodlands, TX). Data for Ca2+-saturated monomeric GCaMP2 were integrated and scaled in HKL2000 (20 ). Data from crystals of Ca2+-dimer, K378W, and 8EF-apo GCaMP2 were processed using Mosflm (21 ) and Scala (22 (link)). Structure Solution, Model Building, and Refinement—All GCaMP2 structures were solved by molecular replacement using the program Phaser (23 ). The Ca2+-saturated dimer structure was solved as described previously (17 ) using the published coordinates of GFP (Protein Data Bank (PDB) entry 1EMA) and the coordinates of M13-bound calmodulin (PDB entry 1CDL) as search models. The G87R Ca2+-bound monomer mutant structure was solved by searching sequentially using the cpEGFP domain and CaM-M13 domains from the refined Ca2+-dimer structure and data between 29.3- and 2.8-Å resolution. Clear solutions were obtained in space group P41212 with translation function Z-scores of 43.1 and 21.3, respectively, for the two domains. Strong positive peaks in the difference map at the expected positions of the calcium ions in CaM (which were omitted from the MR model) indicated the correctness of the solutions. The K378W mutant crystals were isomorphous with those of the G87R mutant and the G87R model was used directly for rigid-body refinement against data from K378W crystals. The Ca2+-saturated monomeric GCaMP2 structure was solved using the refined K378W coordinates as a search model. A clear solution was obtained in space group P21212 with a translation function Z-score of 39.2 using data between 45.4- and 2.65-Å resolution. The 8EF-apo GCaMP2 calcium-free mutant structure was solved by searching for the cpEGFP domain from the Ca2+-dimer structure. A clear solution was obtained in space group C2 with a translation function Z-score of 18.9 using data between 31.9- and 2.8-Å resolution. Subsequently searching for the calcium-free N-terminal or C-terminal lobes of CaM (PDB code 1CFD (24 (link))) did not reveal any clear solutions. Some positive difference density was present in the electron density maps calculated using the cpEGFP domain solution that suggested the position of the N-terminal lobe of CaM, which was placed manually into density and refined. The correctness of this CaM N-terminal lobe placement was indicated by additional positive difference density for the linker connecting cpEGFP and the CaM N-terminal lobe, which was subsequently built. All models were improved by iterative cycles of model building in Coot (25 ) and positional refinement in REFMAC (26 ). Final GCaMP2 models have reasonable R-factors and model geometries, as illustrated in Table 1. A portion of the electron density map for each structure is provided in supplemental Fig. S1.
ALS BL8.2.2, Advanced Light Source Beam Line 8.2.2.
Data collected on home source using Rigaku Rotating Copper Anode RUH3R.
B-factors were calculated on the STAN server using MOLEMAN2 (xray.bmc.uu.se/cgi-bin/gerard/rama_server.pl).
Size Exclusion Chromatography (SEC)—All SEC was carried out using a Superdex 200 10/300 GL column (GE Healthcare) at a flow rate of 0.5 ml min-1 in 20 mm Tris, 100 mm NaCl, 2 mm CaCl2, pH 8.0, for calcium-saturated samples or with 5 mm EGTA in place of CaCl2 for calcium-free samples. Molecular weights were estimated by comparison with elution volumes of standard proteins (Bio-Rad). Sedimentation Velocity Analytical Ultracentrifugation—Analytical ultracentrifugation of GCaMP2 samples was carried out in a Beckman XL-I analytical ultracentrifuge (Beckman Coulter, Fullerton, CA) within the Biophysics Instrumentation Facility at the Massachusetts Institute of Technology. Absorbance scans at 280 nm were collected on calcium-free (24 μm) and calcium-saturated (28 μm) GCaMP2 samples in two-sector cells within a four-hole AnTi-60 rotor at 42,000 rpm. Data were collected at 20 °C in the same buffers as the SEC experiments. Absorbance scans were modeled using a continuous c(s) distribution within Sedfit (27 (link)), correcting for buffer density and viscosity and using a partial specific volume of 0.7300 cm3 g-1. Molecular weight of observed species (Fig. 2B) was estimated based on the best-fit frictional ratio as determined by Sedfit for each sample.
Crystal structures of GCaMP2.A, schematic of the primary amino acid sequence of GCaMP2 illustrating the domain organization. Domains are colored as depicted in B-D. Carets below the schematic show the positions of inter-domain linkers whose amino acid sequences are given. B, stereoview of the structure of the Ca2+-saturated domain-swapped GCaMP2 dimer, depicted as ribbons. One molecule of the dimer is colored by domain as in A, the other molecule is colored light gray. The EGFP chromophore is represented as sticks and calcium ions are shown as orange spheres. C, structure of Ca2+-saturated GCaMP2 monomer, represented as in B except the domains are labeled. D, structure of calcium-free GCaMP2, represented as in B and C. Note that the M13 peptide and the C-terminal half of CaM are not included in the model due to lack of electron density, suggesting their flexibility. This and other structure figures were prepared using PyMOL (Delano Scientific, San Carlos, CA).
Two-photon Laser Scanning Microscopy of Neurons Expressing GCaMP2—To measure intracellular [GCaMP2], acute brain slices containing neurons expressing GCaMP2 were prepared and imaged as previously described (6 (link), 16 (link)). Purified GCaMP2 was diluted into pipette internal solution supplemented with 1 mm K2EGTA at 0.1, 1, and 10 μm concentrations. Each solution was drawn into a thin glass capillary (ID = 0.02 mm, Vitrocom number RT5002). Their fluorescence intensities were measured under two-photon excitation with identical parameters (910 nm excitation) to neuron imaging. 0.1 μm GCaMP2 was not bright enough to significantly exceed PMT dark current at laser powers used for neuronal imaging. Intracellular GCaMP2 concentration in neurons with robust fluorescent responses to action potential firing was estimated by a linear extrapolation from the purified 10 μm GCaMP2 fluorescence intensity. Intracellular GCaMP2 was assumed to be in the apo state (6 (link)). Generation and Screening of GCaMP2 Mutants—Mutants of GCaMP2 were prepared by site-directed mutagenesis (see supplemental Tables S1-S3) and confirmed by sequencing. Preliminary screening for variants with altered oligomerization equilibria (supplemental Table S1) was performed by passing 100-μl aliquots of cell-free extract from 200-ml cultures of overexpressed GCaMP2 over a Superdex 200 10/300 GL column while monitoring the absorption at 280 and 495 nm. Spectrophotometric Analysis—Absorbance spectra were obtained in a Safire2 (Tecan) with UVStar 96-well plates (Greiner) for both the calcium-free (10 mm EGTA) and calcium-loaded state (10 mm CaCl2). For fluorescence spectra, Fluotrac 200 plates (Greiner) were used. Samples were diluted 10-fold in zero free calcium buffer (Invitrogen) (30 mm MOPS, 100 mm KCl, 10 mm EGTA, pH 7.2) for calcium-free spectra, and in 39 μm free calcium buffer (Invitrogen) (30 mm MOPS, 10 mm Ca-EGTA in 100 mm KCl, pH 7.2) for calcium-loaded spectra. For absorbance measurements, samples were dialyzed into 20 mm Tris, 100 mm NaCl, and EGTA or CaCl2 was added to a final concentration of 10 mm.
Akerboom J., Rivera J.D., Guilbe M.M., Malavé E.C., Hernandez H.H., Tian L., Hires S.A., Marvin J.S., Looger L.L, & Schreiter E.R. (2009). Crystal Structures of the GCaMP Calcium Sensor Reveal the Mechanism of Fluorescence Signal Change and Aid Rational Design. The Journal of Biological Chemistry, 284(10), 6455-6464.
The Zn2+-HDAC8 variants were used for crystallographic studies. A new crystal form of HDAC8 was discovered for the HDAC8-TSA and HDAC8-APHA complexes using minor modifications of previously published conditions (18 (link)). The inhibitors TSA and APHA were purchased from Sigma and used without further purification. Briefly, a 4 μL hanging drop of 5 mg/mL HDAC8 in 50 mM Tris (pH 8.0), 150 mM KCl, 5% glycerol, 1 mM dithiothreitol, and 2 mM inhibitor was mixed with a 4 μL drop of precipitant buffer (0.1 M 2-(N-morpholino)ethanesulfonic acid (MES) (pH 5.3), 1–5 % polyethylene glycol (PEG) 6000, 2 mM Tris-2-carboxyethylphosphine (TCEP)) and a 0.4 μL drop of 0.3 M Gly-Gly-Gly, and was equilibrated against a 600 μL reservoir of precipitant buffer at room temperature. The final pH in the crystallization drop was 5.8. Large plate-like crystals appeared within 1 – 5 days and grew to typical dimensions of 300 × 150 × 50 μm3. Crystals were harvested and cryoprotected in 25 mM Tris-HCl, 50 mM MES (pH 5.8), 75 mM KCl, 0.5 mM TCEP, 50 μM inhibitor, and 8% or 20% polyethylene glycol (PEG) 6000 for the APHA and TSA complexes, respectively, or 10% or 30% glycerol for the TSA and APHA complexes, respectively. The D101 HDAC8 variants were similarly crystallized and cryoprotected with slight modification of the precipitant buffer solution. Specifically, the D101A, D101L, D101N, and D101E variants complexed with M344 (purchased from Sigma) crystallized with 1–5% PEG 6000, PEG monomethyl ether 550, PEG 35,000, and PEG dimethyl ether 2,000, respectively. Crystals were cryoprotected in the same buffer with a final glycerol concentration of 30% and 50 μM inhibitor. The H143A variant complexed with an acetylated tetrapeptide substrate (N-acetyl-arginine-histidine-acetyllysine-acetyllysine-coumarin) was crystallized using a precipitant buffer of 50 mM Tris-HCl (pH 8.0), 50 mM MgCl2, 150 mM KCl, 13% PEG 6000, 2 mM TCEP, and 3.2 mM substrate. Crystals were subsequently transferred to a cryoprotectant buffer of 50 mM Tris-HCl (pH 8.0), 25 mM MgCl2, 75 mM KCl, 20% PEG 6000, 1 mM TCEP, 20% glycerol, and 50 μM substrate as described previously for Y306F HDAC8 (25 (link)). Diffraction data were measured on beamline F1 at the Cornell High Energy Synchrotron Source (CHESS, Ithaca, NY) for crystals of the HDAC8-TSA complex, and beamlines XL12-B and X29 at the Brookhaven National Synchrotron Light Source (NSLS, Brookhaven, NY) for crystals of the HDAC8-APHA and D101E HDAC8 variant-M344 complexes. Diffraction data for the remaining D101 HDAC8 variant-M344 complexes and the H143A HDAC8-substrate complex were collected at beamlines ID-24C/E at the Advanced Photon Source, Northeastern Collaborative Access Team (APS, NECAT, Argonne, Il). Crystal and data collection statistics are recorded in Table 1. Data were indexed and merged using HKL2000 (28 ) and MOSFLM (29 ). Molecular replacement calculations were performed with AMoRe (30 ) using the atomic coordinates of an inhibited form of HDAC8 less inhibitor and solvent atoms (PDB accession code 1W22) (18 (link)) as a search probe for rotation and translation functions for the HDAC8-TSA complex. This refined solution was used for molecular replacement with the HDAC8-APHA complex. Similarly, molecular replacement calculations for the D101 HDAC8 variant-M344 complexes and the H143A HDAC8-substrate complex were performed with AMoRe (30 ) or PHASER (31 ) using the structure of wild-type HDAC8 minus ligand and solvent atoms as a search probe (PDB accession codes 1T67 and 2V5W, respectively) (17 (link), 25 (link)). Iterative cycles of refinement and model building were performed using CNS (32 (link)) and O (33 (link)), respectively, in order to improve each structure as guided by Rfree values. Strict noncrystallographic symmetry (NCS) restraints were initially used during the first few cycles of refinement of each complex and relaxed into appropriately-weighted restraints early in refinement. In the HDAC8-TSA and HDAC8-APHA complexes, residues M1–Q12 at the N-termini of all monomers, residues A32-K33 of monomer A, and residues G86-E95 of monomer C of the HDAC8-TSA complex appeared to be disordered and were excluded from the final model. For the HDAC8-APHA complex, residues Q84 to E106 of monomer C appeared to be disordered and were excluded from the final model (APHA was not observed to bind to monomer C). For the HDAC8-D101 variants complexed with M344, disordered regions correspond to those previously identified for the wild-type enzyme (17 (link)). Finally, residues M1-S13 in all monomers of the H143A HDAC8-substrate complex are disordered and excluded from the final model. All refinement statistics are recorded in Table 1.
Dowling D.P., Gantt S.L., Gattis S.G., Fierke C.A, & Christianson D.W. (2008). Structural Studies of Human Histone Deacetylase 8 and its Site-Specific Variants Complexed with Substrate and Inhibitors. Biochemistry, 47(51), 13554-13563.
Nimodipine was a gift sample from Strides Pharma Science limited (India). Cremophor® RH-40 (Polyoxyl 40 hydrogenated castor oil) was procured from Himedia (India). Lipoxol 300 (PEG 300) was procured from Sasol Chemicals (USA). Polyethylene glycol 400 (PEG 400) was obtained from TCI (India). Caprol® ET (hexaglycerol octasterate), Captex® 200 (propylene glycol dicaprylate), Captex® 300 (glyceryl tricaprylate/tricaprate) were gift samples from Abitech (USA). Labrafac™ PG (propylene glycol dicaprylocaprate) from Gattefossé (Canada) was received as a gift sample. Propylene glycol and Ethylene diamine tetra acetic acid disodium (EDTA disodium) were obtained from CDH (India). Chitosan with 90% deacetylation (DA) was acquired from Marine Hydrocolloids (India). Except for the ones we talked about, we used high-quality chemicals for the research. We used them just as they were, without any changes.
Kumar M., Chawla P.A., Faruk A., & Chawla V. (2024). Solid self-nanoemulsifying drug delivery systems of nimodipine: development and evaluation. Future Journal of Pharmaceutical Sciences, 10(1).
HBV was purified from HepAD38 cells using heparin-affinity chromatography as previously reported (95 ). HepG2-NTCP cells were infected with HBV MOI 300 with 4% polyethylene glycol-8000, washed after 6 h and treatments applied.
Harris J.M., Magri A., Faria A.R., Tsukuda S., Balfe P., Wing P.A, & McKeating J.A. (2024). Oxygen-dependent histone lysine demethylase 4 restricts hepatitis B virus replication. The Journal of Biological Chemistry, 300(3), 105724.
A solution of Rapamycin (Selleckchem #S1039) was prepared at the concentration of 2 mg/ml in a mixture of water, 2% dimethylsulfoxide (DMSO), 30% polyethylene glycol 300 (PEG 300), and 5% polysorbate 80 (Tween 80). This solution was administered to the pregnant female via intraperitoneal (i.p.) injection at a dose of 4 mg/kg (twice daily).
Ho H’ng C., Amarasinghe S.L., Zhang B., Chang H., Qu X., Powell D.R, & Rosello-Diez A. (2024). Compensatory growth and recovery of cartilage cytoarchitecture after transient cell death in fetal mouse limbs. Nature Communications, 15, 2940.
A set of samples previously explored to establish the basics of APFD mode was reused here as a control of general operation and to check for reproducibility, and thus, details on most samples as well as reference APFD without TIMS were already communicated [25 (link)–27 (link)]. Solvents of LC–MS grade and polyethylene glycol 300 were obtained from Merck KGaA (Darmstadt, Germany). A sample of triphenylene was available within the author’s institution. The analytes are compiled in Table 1.
Compounds analyzed by APFD-MS in the order of appearance
Paclitaxel was dissolved in DMSO and then diluted in Tween 80, polyethylene glycol 300, ddH2O, and PBS to a final concentration of 2.5 mg/mL. The stock solution was aliquoted and frozen at −20 °C until needed. After determining mouse weight, the stock solution was further diluted in sterile PBS for intraperitoneal injection.
Ortiz A., Stavrou A., Liu S., Chen D., Shen S.S, & Jin C. (2024). NUPR1 packaged in extracellular vesicles promotes murine triple-negative breast cancer in a type 1 interferon-independent manner. Extracellular vesicles and circulating nucleic acids, 5(1), 19-36.
Polyethylene glycol 300 is a water-soluble, non-toxic, and odorless liquid chemical compound. It is commonly used as a solvent, humectant, and plasticizer in various laboratory applications.
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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.
PEG300 is a polyethylene glycol with an average molecular weight of 300 g/mol. It is a clear, colorless, and odorless liquid. PEG300 is commonly used as a solvent, plasticizer, and surfactant in various pharmaceutical, cosmetic, and industrial applications.
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Tween 80 is a non-ionic surfactant and emulsifier. It is a viscous, yellow liquid that is commonly used in laboratory settings to solubilize and stabilize various compounds and formulations.
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The HP 6890 is a gas chromatograph (GC) system designed for high-performance separation and analysis of chemical compounds. It features a programmable oven, adjustable gas flow controls, and advanced data acquisition and processing capabilities. The HP 6890 is a versatile instrument suitable for a wide range of analytical applications in various industries.
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Ethanol is a clear, colorless liquid chemical compound commonly used in laboratory settings. It is a key component in various scientific applications, serving as a solvent, disinfectant, and fuel source. Ethanol has a molecular formula of C2H6O and a range of industrial and research uses.
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Polyethylene glycol is a water-soluble and nontoxic synthetic polymer. It is commonly used as a lubricant, solvent, and dispersing agent in various laboratory and industrial applications.
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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.
Polyethylene glycol 300 (PEG300) is a water-soluble, non-toxic, and non-volatile liquid. It is commonly used as a solvent, emulsifier, and humectant in various pharmaceutical, cosmetic, and industrial applications.
Polyethylene Glycol 300 (PEG 300) is a low molecular weight, water-soluble polymer with a versatile range of applications in biomedical research and pharmaceutical formulations. This nontoxic, odorless, and colorless compound is commonly used as a solvent, carrier, and excipient, owing to its excellent solubilizing and stabilizing properties.
Polyethylene Glycol 300 is particularly useful in the development of drug delivery systems, tissue engineering, and as a cryoprotectant for biomolecules. It is a valuable tool for researchers working in these areas due to its ability to enhance solubility, stability, and preservation of various compounds and biological materials.
PubCompare.ai allows you to screen protocol literature more efficiently and leverage AI to pinpoint critical insights. The platform can help researchers identify the most effective protocols related to Polyethylene Glycol 300 for their specific research goals. The AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuaracy.
Yes, there are various molecular weight versions of Polyethylene Glycol, each with slightly different properties and applications. PEG 300 is just one of the many PEG variants, with a relatively low molecular weight compared to other PEG types. Researchers should consider the specific needs of their project when selecting the most appropriate PEG 300 or alternative PEG formulation.
One key challenge in using Polyethylene Glycol 300 is ensuring optimal solubility, stability, and compatibility with other components in a formulation or application. PubCompare.ai's AI-driven platform can assist researchers by identifying the most effective protocols from the literature, preprints, and patents, helping to navigate these complexities and enhance the reproducibility and accuracy of PEG 300 studies.
More about "Polyethylene glycol 300"
Polyethylene glycol (PEG) 300 is a versatile, low molecular weight, water-soluble polymer with a wide range of applications in biomedical research and pharmaceutical formulations.
This nontoxic, odorless, and colorless compound, also known as Macrogol 300 or PEG-300, is commonly used as a solvent, carrier, and excipient in drug delivery systems, tissue engineering, and cryoprotection for biomolecules.
Its excellent solubilizing and stabilizing properties make it a valuable tool for researchers.
In addition to its uses in biomedical applications, PEG 300 has applications in other areas, such as personal care products, cosmetics, and industrial processes.
It can be combined with other compounds, such as DMSO (Dimethyl Sulfoxide), Tween 80, and Ethanol, to create formulations with enhanced properties.
PEG 300 is particularly useful in the development of drug delivery systems, where it can improve the solubility, stability, and bioavailability of active pharmaceutical ingredients (APIs).
It is also employed in tissue engineering, where it can serve as a scaffold material or a cryoprotectant for preserving cells and biomolecules during freezing and storage.
Researchers can leverage the AI-driven platform of PubCompare.ai to optimize their PEG 300 studies.
The platform helps them locate the best protocols from literature, preprints, and patents, while providing data-driven decisions and enhancing reproducibility and accuracy.
This can lead to more efficient and effective PEG 300 research, ultimately contributing to advancements in biomedical and pharmaceutical fields.
