Ym 30 filter

Manufactured by Merck Group

Sourced in Ireland, United States

The YM-30 filter is a laboratory filtration device manufactured by Merck Group. It is designed to separate and retain molecules or particles larger than 30,000 Daltons from a liquid sample while allowing smaller components to pass through. The core function of the YM-30 filter is to enable sample purification and concentration during various research and analytical processes.

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Lab products found in correlation

Speedvac, by Thermo Fisher Scientific (1 mentions) Trypsin, by Promega (1 mentions) Ultimate 3000 rslcnano hplc system, by Thermo Fisher Scientific (1 mentions) Me220, by Covaris (1 mentions)

Close spelling variants of this product

Microcon filters YM-30 Filter YM-30 (30 kD) Microcon YM-30 centrifugal filter unit Amicon YM-30 filter device Microcon YM-30 filters Microcon YM-30 filter units YM-30

4 protocols using ym 30 filter

Exosomal Protein Extraction and Tryptic Digestion

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Exosomal
protein extraction and tryptic digestion were performed using the
filter-aid sample preparation method. After removing the supernatant,
the exosome pellet was lysed with 30 μL of lysis buffer composed
of 0.1 M Tris-HCl, 4% SDS, and 0.1 M 1,4-dithiothreitol at 99 °C
for 5 min. The extract was cooled down to room temperature, mixed
with 200 μL of 8 M urea, transferred to a centrifugal spin YM-30
filter (Millipore, Billerica, MA), and centrifuged at 14,000g for 20 min. Then, 100 μL of 50 mM iodoacetamide
in 8 M urea was added to the filter and incubated in the dark for
20 min. The filter was then subsequently washed with 8 M urea and
50 mM NH₄HCO₃, respectively, three times each.
Then, 400 ng of trypsin (Promega, Madison, MI) in 75 μL of 50
mM NH₄HCO₃ was added to the filter and incubated
at 37 °C overnight. The released peptides were collected by centrifugation
and desalted with a ZipTip C18 tip (Millipore, Billerica, MA). The
eluted peptides were dried down in a SpeedVac concentrator (Thermo)
for the following mass spectrometry analysis.

Zhu J., Tan Z., Zhang J., An M., Khaykin V.M., Cuneo K.C., Parikh N.D, & Lubman D.M. (2022). Sequential Method for Analysis of CTCs and Exosomes from the Same Sample of Patient Blood. ACS Omega, 7(42), 37581-37588.

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Titin Identification via SDS-PAGE and Mass Spectrometry

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The presence of titin in the sample was confirmed by SDS-PAGE (Supplementary Figure S1) and mass spectrometry analysis (Supplementary Table S1). SDS-PAGE of titin was performed as in [71 (link)] with our modification. In particular, our experiments used a separating gel containing 6.5–7% polyacrylamide prepared as described in [71 (link)]. For shotgun mass spectrometry analysis the sample was solubilized into a buffer (4% sodium dodecyl sulfate in 0.1 M Tris-HCl pH 7.6, 0.1 M dithiothreitol) and incubated for 5 min at 95 °C as described [72 ]. The samples were sonicated (4 × 30 s at 20 W; ME220, Covaris, Woburn, MA, USA), centrifuged (5 min, 16,000× g), and the supernatant was collected. The YM-30 filter (Millipore, Ireland) was used for alkylation and trypsinolysis (14 h, 2 μg of trypsin (Tripsin Gold, Promega, Madison, WI, USA)) according to the FASP method [73 ]. Peptides were desalted using C18 microcolumns and subjected to HPLC-MS/MS analysis using the HPLC Ultimate 3000 RSLCnano system (Thermo Scientific, Waltham, MA, USA) as described [72 ]. For a list of detected titin peptides, see Supplementary Table S1.

Bobylev A.G., Fadeev R.S., Bobyleva L.G., Kobyakova M.I., Shlyapnikov Y.M., Popov D.V, & Vikhlyantsev I.M. (2021). Amyloid Aggregates of Smooth-Muscle Titin Impair Cell Adhesion. International Journal of Molecular Sciences, 22(9), 4579.

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Oligonucleotide Library Generation and Selection

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A library of random ssDNA oligonucleotides was from BD Biosciences. This library consisted of a 30 base random region with a flanker of a 21 base 5′-primer (5′-CGCTCGATCGATCTAGATTCG-3′) and a 21 base 3′-primer (5′-GTCATCACGCTCAGATCACTG-3′). PCR amplification was repeated using the extracted ssDNA oligonucleotides after each selection round. First, the ssDNA random library was incubated with Taq DNA polymerase in a mixture of dATP, dTTP, dCTP, and dGTP (500 μM each) for 5 hours at 37°C. After reverse strand was synthesized, the dsDNA library (400 nM) went through PCR amplification. Then PCR products were filtered by a Millipore YM-30 filter to purify dsDNA product. The product was incubated with streptavidin-coated magnetic beads in buffer (10 mM Tris–HCl, 2 M NaCl, and 1 mM EDTA [pH 7.5]) for 20 minutes. Biotin on the reverse strand of the dsDNA binds to the streptavidin beads, followed by separation of the two DNA strands in melting solution (0.1 M NaOH). The forward strands were collected after filter with Millipore YM-10 filter for the next round of selection.

Chen L., He W., Jiang H., Wu L., Xiong W., Li B., Zhou Z, & Qian Y. (2018). In vivo SELEX of bone targeting aptamer in prostate cancer bone metastasis model. International Journal of Nanomedicine, 14, 149-159.

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Depletion of H-ficolin and L-ficolin

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Cibacron Blue agarose was used to co-deplete H-ficolin and human serum albumin. Briefly, 0.1 ml of positive or negative human serum was mixed with an equal volume of PBS, loaded on rehydrated SwellGel Disc (ThermoFisher Scientific, Waltham, MA, USA) in a spin column, and incubated at room temperature for 5 min. The supernatant was collected and re-applied to the column twice to derive H-ficolin/albumin-depleted sera. The bound albumin and the associated H-ficolin were eluted with PBS + 500 mM NaCl, dialyzed against PBS, and concentrated to 0.2 ml using YM-30 filter (Millipore, Billerica, MA, USA). To deplete L-ficolin, 0.1 mg of monoclonal antibody against L-ficolin (R&D Systems, Minneapolis, MN, USA) was immobilized on 0.1 ml of SiMAG/MP-COOH magnetic beads (Chemicell, Berlin, Germany) using EDC and NHS (Pierce, Rockford, IL, USA) following the manufacturer's instruction. Briefly, 0.1 ml of positive human serum were diluted four times with PBS, mixed with L-ficolin-immobilized magnetic beads, and incubated at room temperature for 2 hrs in tubes. The supernatants containing the depleted sera were recovered by placing the tubes in the magnetic rack. The bound ficolins were eluted with 0.4 ml of 100 mM glycine (pH 2.8), neutralized with 0.1 ml of 1 M Tris-HCl (pH 8.0), dialyzed against PBS, and concentrated to 0.2 ml using a YM-30 filter.

Lei X., Liu C., Azadzoi K., Li C., Lu F., Xiang A., Sun J., Guo Y., Zhao Q., Yan Z, & Yang J. (2015). A novel IgM–H-Ficolin complement pathway to attack allogenic cancer cells in vitro. Scientific Reports, 5, 7824.

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