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3 kda mwco

Manufactured by Merck Group
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The 3 kDa MWCO (Molecular Weight Cut-Off) is a lab equipment product used for separating and purifying molecules based on their size. It is designed to retain molecules with a molecular weight greater than 3 kilodaltons (kDa) while allowing smaller molecules to pass through. This product is commonly used in various applications, such as protein purification, sample preparation, and dialysis.

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Lab products found in correlation

0.22 μm filter, by Merck Group (1 mentions) Synergy h1 multi mode reader, by Agilent Technologies (1 mentions) Zetasizer nano zs instrument, by Malvern Panalytical (1 mentions) Whatman, by Cytiva (1 mentions) 815 cd spectropolarimeter, by Jasco (1 mentions) Spectra manager, by Jasco (1 mentions) Bradford assay, by Thermo Fisher Scientific (1 mentions) Penicillin and streptomycin, by Solarbio (1 mentions) Galaxy 170, by Eppendorf (1 mentions) Fetal bovine serum (fbs), by Clark Bioscience (1 mentions) L dmem, by Solarbio (1 mentions) Fetal bovine serum (fbs), by Solarbio (1 mentions) L dmem, by Clark Bioscience (1 mentions) L glutamine, by Solarbio (1 mentions) Microplate reader, by Tecan (1 mentions) Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions)

Spelling variants of this product

3 kDa MWCO filters (6 citations) Amicon 3,000 KDa MWCO filtration unit (3 citations) Amicon Ultra (3 kDa MWCO) (2 citations) Amicon 3 kDa MWCO centrifugal filters (2 citations)

8 protocols using 3 kda mwco

1

Tumor Cell Secretome Isolation

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The normal medium was removed from 80% confluent cells and cells were washed three times with PBS before addition of serum-free medium. Tumor cells were cultured in serum-free medium for 2 days. Then the medium was collected and centrifuged at 3000×g for 10 min to remove cell pellet. The medium was fractioned at 3 KDa MWCO (Millipore) at 4000×g for 50 min and filtered by passing through a 0.22μm filter (Millipore) before use or stored at -80°C.
Tian T., Li X., Hua Z., Ma J., Wu X., Liu Z., Chen H, & Cui Z. (2017). S100A7 promotes the migration, invasion and metastasis of human cervical cancer cells through epithelial–mesenchymal transition. Oncotarget, 8(15), 24964-24977.
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2

Micellar Nanoparticle Formulation and Characterization

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Self-assembled micellar nanoparticles (NP) were obtained by first dissolving lyophilized polymer at 50 mg/ml in 100% ethanol, then rapidly pipetting dissolved polymer into 100 mM phosphate buffer (pH 7) to a final concentration of 10 mg/ml. Nanoparticles were formulated in the same way for both pH-responsive (NPpH) and control (NPctrl) polymers. For in vivo studies, ethanol was removed by buffer exchange into PBS (pH 7.4) via 3 cycles of centrifugal dialysis (Amicon, 3 kDa MWCO, Millipore), and NP solutions were then sterilized via syringe filtration (Whatman, 0.22 μm, GE Healthcare). Final polymer concentration was determined with UV-Vis spectrometry (Synergy H1 Multi-Mode Reader, BioTek) by measuring absorbance of aromatic PDS groups at 280 nm. Size of the NP was measured via dynamic light scattering (DLS). NP solutions were prepared at a concentration of 0.1–0.2 mg/ml in PBS (pH 7.4) and the hydrodynamic radius was measured using a Malvern Instruments Zetasizer Nano ZS Instrument (Malvern, USA). Representative DLS data for both polymers at physiological pH (7.4) can be found in Figure S1E. In addition, size change of NPpH but not NPctrl at pH 5.8, as measured by DLS, can be seen in Figure S1F (left).
Knight F.C., Gilchuk P., Kumar A., Becker K.W., Sevimli S., Jacobson M.E., Suryadevara N., Wang-Bishop L., Boyd K.L., Crowe JE J.r., Joyce S, & Wilson J.T. (2019). Mucosal Immunization with a pH-Responsive Nanoparticle Vaccine Induces Protective CD8+ Lung-Resident Memory T Cells. ACS nano, 13(10), 10939-10960.
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3

Circular Dichroism Analysis of Protein

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Protein samples were extensively dialyzed into 20 mM NaPhosphate, 20 mM NaCl pH7.5 and then concentrated to 0.2 mg/ml in spin concentrators (0.5 ml, 3KDa MWCO, Millipore). The Circular dichroism (CD) analysis was done on a JASCO 815 CD spectropolarimeter. Data are averages of 5 independent scans in the 190 nm –250 nm range, and were normalized to the baseline of the dialysis buffer. The data were smoothed using the manufacturer’s software (Jasco SpectraManager) before interpretation. The percentage of α-helix was calculated according to the formula: percentage of α-helix = (θ208–4000)/(-33000-4000)×100, where θ208 is the ellipticity at 208 nm [103] (link).
Lo M.K., Søgaard T.M, & Karlin D.G. (2014). Evolution and Structural Organization of the C Proteins of Paramyxovirinae. PLoS ONE, 9(2), e90003.
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4

Serum-free Conditioned Media Isolation

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Cells were grown to 70–80% confluence, rinsed twice in PBS and placed in serum-free media for 48 h. CM was cleared by centrifugation (1500g × 5 min, 4°C), the supernatants concentrated by spin-filtration (3-kDa MWCO, Millipore, Billerica, MA) and stored at −80°C. Protein concentration was measured using Bradford Assay (Thermo Scientific, Waltham, MA).
Nwani N.G., Deguiz M.L., Jimenez B., Vinokour E., Dubrovskyi O., Ugolkov A., Mazar A.P, & Volpert O.V. (2016). Melanoma cells block PEDF production in fibroblasts to induce the tumor-promoting phenotype of cancer-associated fibroblasts. Cancer research, 76(8), 2265-2276.
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5

Isolation and Characterization of ADSCs

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Adipose tissues were obtained from the subcutaneous abdominal fat and digested with collagenase I at 37°C for 45 min with continuous shaking. After neutralizing enzyme activity with L-DMEM (low glucose-Dulbecco's modified Eagle medium) supplemented with 10% FBS (Clark, USA), the homogenate was filtered and centrifuged, and the ADSCs were suspended in L-DMEM supplemented with 10% FBS, 2 mM L-glutamine, and 100 μg/ml penicillin and streptomycin (Solarbio, China). The cells were cultured at 37°C under 5% CO2 in a humidified incubator (Galaxy 170 S, Eppendorf, Germany). The ADSCs were characterized as previously described [31 (link)], cultured in serum-free L-DMEM for 48 h. Then, the medium was aspirated and centrifuged at 1000 g for 15 min at 4°C to remove the cell debris. The supernatant was then centrifuged at 5000 g for 50 min at 4°C using 3 kDa MWCO (Millipore, Billerica, USA) to concentrate it by 25-fold. The CM aliquots were transferred to sterile 1.5 ml EP tubes and stored at -80°C.
Jiao Z., Ma Y., Wang Y., Liu T., Zhang Q., Liu X., Piao C., Liu B, & Wang H. (2021). Protective Effect of Adipose-Derived Mesenchymal Stem Cell Secretome against Hepatocyte Apoptosis Induced by Liver Ischemia-Reperfusion with Partial Hepatectomy Injury. Stem Cells International, 2021, 9969372.
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6

Quantifying Immobilized Cadherin Density

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Hydrogels functionalized with fluorescein (FITC)-labeled N-Cad-Fc were used to measure the density of immobilized cadherin (molecular number/area). The FITC-labeled N-Cad-Fc was prepared by reacting 1 mg/mL of N-Cad-Fc dissolved in HEPES buffer with fluorescein isothiocyanate (Sigma-Aldrich) for 12 h at 4 °C. Following the reaction, FITC-labeled N-Cad-Fc was purified using centrifugal filtration (3 kDa MWCO, Millipore), followed by dialysis in HEPES buffer at 4 °C to remove unreacted FITC. While FITC is reactive towards amine groups on the protein, it most stably binds to the N-terminus19 , which ensures that the protein is mostly unaltered, and thus minimally alters the immobilization of the protein onto the hydrogel. Then, the FITC-labeled N-Cad-Fc was conjugated to a hydrogel in calcium containing HEPES buffer. The concentration of FITC-N-Cad-Fc was varied from 0 to 250 µg/mL. Then, the gel was washed three times with fresh PBS to remove excess FITC-N-Cad-Fc. The fluorescence from the gels was measured using the microplate reader (Tecan). Three gels per condition were analyzed. Separately, fluorescence yield of the HEPES buffer dissolved with known amounts of FITC-labeled N-Cad-Fc were measured to prepare a standard curve. This standard curve was used to back-calculate the density of N-Cad-Fc immobilized within the gel.
Vega L. J.C., Lee M.K., Qin E.C., Rich M., Lee K.Y., Kim D.H., Chung H.J., Leckband D.E, & Kong H. (2016). Three Dimensional Conjugation of Recombinant N-Cadherin to a Hydrogel for In Vitro Anisotropic Neural Growth. Journal of materials chemistry. B, Materials for biology and medicine, 4(42), 6803-6811.
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7

Fractionation of Oligosaccharides and Proteins

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The BCF (4 mg/mL) was fractionated by centrifugal filtration using a 3 kDa MWCO (Merck Millipore, Cork, Ireland). HPAEC-PAD analysis, as described above, confirmed the permeate contained the oligosaccharides and lactose while the retentate contained proteins and peptides greater than 3 kDa (confirmed by HPLC, previously described above). The permeate was examined in the adhesion assay described below.
Morrin S.T., McCarthy G., Kennedy D., Marotta M., Irwin J.A, & Hickey R.M. (2020). Immunoglobulin G from bovine milk primes intestinal epithelial cells for increased colonization of bifidobacteria. AMB Express, 10, 114.
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8

Quantification of Inflammatory Cytokines in Bone Marrow and Serum

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Human proteins IL-8, TNFα, IL-6, IL-1β, IL-27, and IL-10 were quantiatedby using a U-plex, multiplex assays, Biomarker Group 1 (Human; MSD, Rockville, MD, United States) in serum samples and bone marrow aspirates of huBRGSF collected at day 3 post-surgery. Cytokines were also measured in supernatants obtained after centrifugation of isolated bone marrow cells in HBSS. Bone marrow samples were concentrated (~10x) using the Amicon Ultra-0.5 centrifugal filter unit with 3KDa MWCO (Merck).
Hofstee M.I., Siverino C., Saito M., Meghwani H., Tapia-Dean J., Arveladze S., Hildebrand M., Rangel-Moreno J., Riool M., Zeiter S., Zaat S.A., Moriarty T.F, & Muthukrishnan G. (2024). Staphylococcus aureus Panton-Valentine Leukocidin worsens acute implant-associated osteomyelitis in humanized BRGSF mice. JBMR Plus, 8(2), ziad005.
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