Amicon centrifugal filter

Manufactured by Merck Group

Sourced in United States, Germany, France, United Kingdom

Amicon centrifugal filters are laboratory equipment designed for the separation and concentration of macromolecules, such as proteins, enzymes, and nucleic acids, from complex solutions. These filters utilize centrifugal force to facilitate the filtration process, allowing for the efficient recovery of the desired components while removing unwanted substances.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

50K Amicon® Ultra Centrifugal Filters 100K Amicon® Ultra Centrifugal Filters Amicon® Ultra 15 mL Centrifugal 3 kDa Filters Amicon® Ultra Centrifugal Filters Ultracell®—3 kDa, Amicon Ultra Centrifugal Filters 100 kb Amicon Ultra 100 K centrifugal filters Amicon ultra-15 centrifugal filters ultracel-3 K Amicon Ultra 3K MWCO Centrifugal Filters Amicon Ultra‐2 Centrifugal Filters Amicon 100 kDa MWCO Ultra Centrifugal filters

166 protocols using amicon centrifugal filter

Purification of Recombinant PsOep23 Protein

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

PsOep23 inclusion bodies were solubilized in 50 mM Tris–HCl pH 8.0, 100 mM NaCl, 8 M urea for 1 h at room temperature. Insoluble material was removed by centrifugation at 20,000 g for 15 min and supernatant containing PsOep23 (equiv. to 150 μg protein) was further purified using a 5 ml HisTrap HP column (GE Healthcare, Germany). Bound PsOep23 was washed with five column volumes (CV) of 50 mM Tris–HCl pH 8.0, 100 mM NaCl, 8 M urea buffer containing 35 mM imidazole. Elution was performed by raising the imidazole concentration to 500 mM.
Fractions enriched in PsOep23 were diluted 20-fold in 100 mM Tris–HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100 and further purified using Strep-Tactin affinity matrix. In brief, the protein was allowed to bind to Strep-Tactin Sepharose (iba, Germany) overnight at 4°C followed by washing with 100 mM Tris–HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.1% CHAPS. Protein elution was achieved by 2.5 mM desthiobiotin (iba) at 4°C for 30 min.
In a second approach nickel affinity purified PsOep23 was subjected to buffer exchange in 20 mM sodium phosphate pH 7.0, 8 M urea using Amicon centrifugal filters (MWCO 10 kDa, Merck). The PsOep23 sample was loaded onto a HiTrap Q FF column (GE Healthcare) and flow-through was collected.

Goetze T.A., Patil M., Jeshen I., Bölter B., Grahl S, & Soll J. (2015). Oep23 forms an ion channel in the chloroplast outer envelope. BMC Plant Biology, 15, 47.

+ Open protocol

+ Expand

Purification of Soluble HIV-1 Envelope Protein

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

FreeStyle 293F cells were transfected with 80 µg of expression plasmids encoding for soluble pConBgp120-His in sterile disposable PETG flasks (Wagner and Munz GmbH, Munich, Germany) with 3 µg polyethylenimine (Sigma Aldrich, Taufkirchen, Germany) per 1 µg DNA. The transfection mix was prepared in OPTI-MEM Reduced Medium (Thermo Fisher, Schwerte, Germany). Medium was changed 6 h after transfection. Three days post-transfection, supernatants were collected and sterile-filtered through 0.2 µm Minisart filters (Sigma Aldrich, Taufkirchen, Germany) and purified via lectin affinity chromatography using lectin from Galanthus nivalis (Vector Laboratories Inc., Burlingame, CA, USA). Columns were loaded after washing with PBS containing 1 mM EDTA and 1 mM EGTA (both Sigma Aldrich, Taufkirchen, Germany). After loading, columns were washed and protein eluted using a 19.5% solution of Methy-α-d-mannopyranosid (Merck, Darmstadt, Germany). Carbohydrates in the eluate were dialyzed. The purified protein was concentrated over Amicon Centrifugal Filters with 10 kDa cut-off (Merck, Darmstadt, Germany). Protein concentration was measured using the ND100-NanoDrop^® (peQlab, Erlangen, Germany). Samples were stored at 4 °C until further use.

Tannig P., Peter A.S., Lapuente D., Klessing S., Damm D., Tenbusch M., Überla K, & Temchura V. (2020). Modulation of Vaccine-Induced HIV-1-Specific Immune Responses by Co-Electroporation of PD-L1 Encoding DNA. Vaccines, 8(1), 27.

+ Open protocol

+ Expand

Purification and Characterization of Recombinant Collagen IV

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

HEK293 cells were grown in DMEM/F12 medium (Sigma-Aldrich) containing 5% FBS and 50 µg/ml ascorbic acid phosphate (Wako Pure Chemical Industries). Transfections were performed by using the calcium phosphate precipitation method and 5 µg of each plasmid DNA individually. Selection of transfected cells was started with 250 µg/ml G418 (Corning) 2 d after transfection. Resistant clones were isolated and expanded. Expression was evaluated using anti-FLAG (Sigma-Aldrich), α1 (IV) NC1, and α2 (IV) NC1-specific antibodies (H11 and H22, respectively; Sado et al., 1995 (link)).
Recombinant proteins were initially purified by affinity chromatography on anti-FLAG M2 affinity agarose (Sigma-Aldrich) according to the manufacturer’s instructions and were further purified by SEC. Recombinant products were concentrated separately using Amicon centrifugal filters (EMD Millipore). Protein concentration was determined with a Nanodrop 2000c spectrophotometer (Thermo Fisher Scientific) using the following conversion factors calculated from PROTPARAM: 1.08 A₂₈₀ = 1 mg/ml for α1-84/α1-CB3 and 1.219 A₂₈₀ = 1 mg/ml for α2-84/α2-CB3.

Cummings C.F., Pedchenko V., Brown K.L., Colon S., Rafi M., Jones-Paris C., Pokydeshava E., Liu M., Pastor-Pareja J.C., Stothers C., Ero-Tolliver I.A., McCall A.S., Vanacore R., Bhave G., Santoro S., Blackwell T.S., Zent R., Pozzi A, & Hudson B.G. (2016). Extracellular chloride signals collagen IV network assembly during basement membrane formation. The Journal of Cell Biology, 213(4), 479-494.

+ Open protocol

+ Expand

Lactate and ATP Quantification Protocol

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

For lactate measurements, medium was sampled from cells 1h and 4h after treatment and deproteinized with Amicon centrifugal filters (Merck Millipore) for lactate dehydrogenase removal. Colorimetric method was performed with lactate assay kit (Sigma-Aldrich) following the manufacturer's instructions. Absorbance was read at 570 nm.
For ATP measurements, cells were lysed with digitonin-based buffer (Cayman). Intracellular ATP was determined by a luciferin/luciferase method using an ATP bioluminescent assay kit (Sigma-Aldrich). Luminescence was measured using a 96-well plate luminometer. Cytosolic ATP content was calculated by an ATP standard curve and normalized to cellular protein content/well.

Leguisamo N.M., Gloria H.C., Kalil A.N., Martins T.V., Azambuja D.B., Meira L.B, & Saffi J. (2017). Base excision repair imbalance in colorectal cancer has prognostic value and modulates response to chemotherapy. Oncotarget, 8(33), 54199-54214.

+ Open protocol

+ Expand

Extracellular Vesicle Purification Protocol

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

PFP (800 μl) was ultracentrifuged at 100,000g for 2 hours at 4 °C to obtain the crude EV pellet. The crude EV pellet was then reconstituted with 500 μl of 0.1μm filtered phosphate buffered saline (PBS) and passed through size exclusion chromatography columns (qEV original, Izon, Medford, MA) for purification according to manufacturer’s instructions. EV numbers and plasma protein levels of each elution fraction were determined by nanoparticle tracking analysis (NTA) and Bradford assay, respectively. EV rich, plasma protein poor fractions (fraction 8–12) were pooled, centrifuged, and concentrated by using 10 kDa Amicon centrifugal filters (Cat # UFC801024, Merck Millipore Ltd.) to obtain pure EVs. Pure EV suspensions in 0.1 μm filtered PBS were then stored in 100 μL aliquots and stored at −80 °C for future analysis.

Yang X., Chatterjee V., Zheng E., Reynolds A., Ma Y., Villalba N., Tran T., Jung M., Smith D.J., Wu M.H, & Yuan S.Y. (2022). Burn injury-induced extracellular vesicle production and characteristics. Shock (Augusta, Ga.), 57(6), 228-242.

+ Open protocol

+ Expand

Proteomic Sample Preparation Procedure

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

DSSO, Dulbecco’s modified Eagle’s medium (DMEM), foetal bovine serum (FBS), l-glutamine, penicillin, streptomycin and phosphate-buffered saline (PBS) were obtained from Thermo Fisher Scientific (Les Ulis, France). Formic acid (FA), HPLC grade water, trifluoroacetic acid (TFA), acetonitrile (ACN), methanol, ethanol, acetone and trichloroacetic acid were all purchased from Biosolve (Dieuze, France). dl-Dithiothreitol (DTT), iodoacetamide (IAA), chloroform, dimethyl sulfoxide (DMSO), ammonium bicarbonate (AB), 4-(2-hydroxyethyl)piperazine-1-ethane sulfonic acid, N-(2-hydroxyethyl)piperazine-N-(2-ethane sulfonic acid) (HEPES), sodium chloride (NaCl) and magnesium chloride (MgCl₂) were obtained from Sigma-Aldrich. Tris was purchased from Bio-Rad (Steenvoorde, France). Extraction Illustra triplePrep Kit was from GE Healthcare. LysC/trypsin was obtained from Promega (Charbonnières-les-Bains, France). Amicon centrifugal filters and C18 ZipTip pipette tips were from Merck Millipore (Merck KGaA, Darmstadt, Germany).

Cardon T., Franck J., Coyaud E., Laurent E.M., Damato M., Maffia M., Vergara D., Fournier I, & Salzet M. (2020). Alternative proteins are functional regulators in cell reprogramming by PKA activation. Nucleic Acids Research, 48(14), 7864-7882.

+ Open protocol

+ Expand

Extracellular Vesicle Isolation from HUVEC

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

HUVECtert cells were treated with OxPAPC (25 µg/mL) in serum-free M199 medium in 12-well plates. After 24 h, the conditioned medium (CM) of three single wells from control cells and OxPAPC-treated cells was pooled, respectively. After centrifugation at 17,000× g for 20 min, supernatants were diluted with PBS to a final volume of 12 mL and centrifuged at 100,000× g for 60 min. Supernatants (10 mL) were concentrated and rebuffered to 50 mM NH₄HCO₃ using Amicon centrifugal filters (Merck KGaA, Darmstadt, Germany) with an MWCO of 3 kDa (final volume: 120 µL). Protein concentration was determined with the MicroBCA Protein Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA).

Mauerhofer C., Afonyushkin T., Oskolkova O.V., Hellauer K., Gesslbauer B., Schmerda J., Ke Y., Zimmer A., Birukova A.A., Birukov K.G, & Bochkov V. (2022). Low Concentrations of Oxidized Phospholipids Increase Stress Tolerance of Endothelial Cells. Antioxidants, 11(9), 1741.

+ Open protocol

+ Expand

HA-TM Protein Production via Lectin Affinity

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

FreeStyle 293F cells were transfected with 80 µg of expression plasmids encoding for soluble HA without transmembrane domain (HA-TM) in sterile disposable PETG flasks (Wagner and Munz GmbH, Munich, Germany) with 3 µg polyethylenimine (Sigma Aldrich, Taufkirchen, Germany) per 1 µg DNA in OPTI-MEM Reduced Medium (Thermo Fisher, Schwerte, Germany). Culture medium was exchanged six hours after transfection. Three days post-transfection, supernatants were collected and sterile filtered through 0.2-µm Minisart filters (Sigma Aldrich, Taufkirchen, Germany) before purification via Erythrina cristagalli (Vector Laboratories Inc., Burlingame, CA, USA) lectin affinity chromatography. After washing with PBS containing 1 mM EDTA and 1 mM EGTA (Sigma Aldrich, Taufkirchen, Germany), columns were loaded with the filtered supernatant. Columns were washed and protein eluted using 200 mM lactose (Sigma Aldrich, Taufkirchen, Germany). Protein samples were concentrated and elution carbohydrates in the eluate dialyzed via Amicon Centrifugal Filters with 10 kDa cut-off (Merck, Darmstadt, Germany). Protein concentration was analyzed using the ND100-NanoDrop^® (peQlab, Erlangen, Germany). Samples were stored at 4 °C until further use. Protein production was monitored with Western Blot and purity assessed by Silver staining (Supplementary Figure S1C).

Tannig P., Peter A.S., Lapuente D., Klessing S., Schmidt A., Damm D., Tenbusch M., Überla K, & Temchura V. (2020). Genetic Co-Administration of Soluble PD-1 Ectodomains Modifies Immune Responses against Influenza A Virus Induced by DNA Vaccination. Vaccines, 8(4), 570.

+ Open protocol

+ Expand

Sortase A-mediated Ligation and Characterization of 1NOG-Antibody Complexes

Cited 1 time

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

The expression and purification procedures of Sortase A and 1NOG-His trimer were described previously (38 (link)). Briefly, the proteins were produced in Rosetta2 E. coli cells (Novagen) and purified with Ni NTA-agarose beads (Qiagen) and Superdex 200 increase 10/300 column (GE Healthcare). For protein ligation reaction, GPCysRRLL-LPETG-Strep and GGGGGSGSP-1NOG-His trimer were mixed with a molar ratio of 3:1 in TBS buffer plus 2 mM CaCl₂ and 1 μM purified Sortase A. After incubation at room temperature for 50 minutes, 2 mM EGTA (final concentration) was added, and the reaction mixture was immediately purified with a Superdex 200 increase 10/300 column (GE Healthcare) to obtain GPCysRRLL-LPETG-1NOG trimer.
To make samples of GPCysRRLL-LPETG-1NOG in complex with antibodies, 5 μg of Sortase A-ligated GPCysRRLL-LPETG-1NOG trimer was incubated with 20 μg of 8.9F-scFv, 12.1F-scFv, 37.2D-scFv, 25.10C-Fab, 19.7E-Fab, or 37.7H-Fab at room temperature overnight. Excess scFv or Fab was removed using 100 KDa cut-off Amicon centrifugal filters (EMD Millipore). The concentrated complexes were further characterized by electron microscope analysis.

Eis-Hübinger A.M., Däumer M., Matz B, & Schneweis K.E. (1999). Evaluation of Three Glycoprotein G2-Based Enzyme Immunoassays for Detection of Antibodies to Herpes Simplex Virus Type 2 in Human Sera. Journal of Clinical Microbiology, 37(5), 1242-1246.

+ Open protocol

+ Expand

Monoclonal Antibody Characterization

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

Four homologous monoclonal antibodies from human myeloma, including IgG1κ, IgG2κ, IgG3κ, and IgG4κ, were purchased from Sigma Aldrich (St. Louis, MO, USA). Ammonium acetate (AA) and cesium iodide (CsI) were purchased from Sigma Aldrich (St. Louis, MO, USA). Borosilicate capillaries with inner and outer diameters of 0.58 mm and 1.0 mm, respectively, were purchased from Sutter Instruments (Novato, CA, USA). Amicon centrifugal filters with a 100 kDa cutoff were purchased from Merck Millipore (Darmstadt, Germany).

Yin Z., Du M., Chen D., Zhang W., Huang W., Wu X, & Yan S. (2021). Rapid structural discrimination of IgG antibodies by multicharge-state collision-induced unfolding. RSC Advances, 11(58), 36502-36510.

+ 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!