pierce 9k mwco protein concentrators  (Thermo Fisher)


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    Pierce 9K MWCO Protein Concentrators
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    87748
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    Thermo Fisher pierce 9k mwco protein concentrators

    https://www.bioz.com/result/pierce 9k mwco protein concentrators/product/Thermo Fisher
    Average 99 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    pierce 9k mwco protein concentrators - by Bioz Stars, 2020-04
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    Clone Assay:

    Article Title: Phosphorylation of Synaptic GTPase-activating Protein (synGAP) by Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII) and Cyclin-dependent Kinase 5 (CDK5) Alters the Ratio of Its GAP Activity toward Ras and Rap GTPases *
    Article Snippet: Paragraph title: Cloning, Expression, and Purification of r-synGAP ... Fractions containing synGAP were pooled and concentrated by ultrafiltration through a 150-kDa cutoff filter (Thermo Scientific, catalog no. 89923) for r-synGAP, and a 9-kDa cutoff filter (Thermo Scientific, catalog no. 89885A) for sr-synGAP.

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: Soluble FcRn expression was assessed by anti-tag enzyme-linked immunosorbent assays (ELISA) and the 8 highest expressing CHO cell clones during early growth were pooled and scaled. .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: The 1,016 base pair product was cloned into pET15b (Novagen‐Merck Millipore), and recombinant protein was obtained by Ni‐NTA agarose column protein purification (Qiagen). .. Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH).

    Centrifugation:

    Article Title: Novel Immunomodulators from Hard Ticks Selectively Reprogramme Human Dendritic Cell Responses
    Article Snippet: Protein purification The medium was cleared by centrifugation (2000 g, 10 min) 72 hours after infection, and proteins were precipitated by adding polyethylene glycol (PEG4000, Sigma; 18 g/100 ml). .. The protein-containing eluate fractions were pooled, concentrated using a 9K MWCO Pierce Protein Concentrator (Thermo Scientific), then further purified by size exclusion chromatography with a Superdex 75 HR10/30 column (GE Healthcare) using PBS (pH7.4) as running buffer.

    Article Title: Phosphorylation of Synaptic GTPase-activating Protein (synGAP) by Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII) and Cyclin-dependent Kinase 5 (CDK5) Alters the Ratio of Its GAP Activity toward Ras and Rap GTPases *
    Article Snippet: Cells were lysed with an ML-110 microfluidizer (Microfluidics), and insoluble material was removed by centrifugation at 30,000 × g . r-synGAP in the supernatant was bound to 1.5 ml of Talon metal affinity resin per g of cell pellet (Clontech, catalog no. 635503) by batch absorption for 90 min. .. Fractions containing synGAP were pooled and concentrated by ultrafiltration through a 150-kDa cutoff filter (Thermo Scientific, catalog no. 89923) for r-synGAP, and a 9-kDa cutoff filter (Thermo Scientific, catalog no. 89885A) for sr-synGAP.

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: After centrifugation of the agarose, the supernatant was discarded and washing steps were performed with NPI-20 wash buffer (50 mM NaH2 PO4 ; 300 mM NaCl; 10 mM imidazole; pH 8). .. Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS.

    Article Title: Isolation and Characterization of the Herpes Simplex Virus 1 Terminase Complex
    Article Snippet: .. The eluted protein sample was concentrated 10-fold by column centrifugation (Pierce; catalog no. 89884A). .. The concentrated proteins were separated by centrifugation on 2.5 to 20% sucrose (in 1× PBS) gradients (SW50.1 rotor at 35,000 rpm for 18 h).

    Article Title: The BH4 Domain of Anti-apoptotic Bcl-XL, but Not That of the Related Bcl-2, Limits the Voltage-dependent Anion Channel 1 (VDAC1)-mediated Transfer of Pro-apoptotic Ca2+ Signals to Mitochondria *
    Article Snippet: The homogenate was purified by centrifugation at 95,000 × g for 30 min on a PercollTM density gradient in isolation buffer 1 (a 15% Percoll gradient on a 30% layer). .. MAMs were centrifuged for 10 min at 7000 × g three times to remove mitochondrial contamination and finally centrifuged at 100000 × g for 1 h. The MAM fraction pellet was collected and concentrated using Pierce Protein Concentrators, 9K MWCO (Thermo Fisher Scientific Inc.) before Western blotting.

    Luciferase:

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: Cell surface Gaussia Luciferase binding assays were performed as previously described ( ). .. After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific).

    Filtration:

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: All IgGs were produced by transient transfection of HEK-freestyle cells (Thermo Fisher Scientific), as previously described by Vink et al. and Dekkers et al. After 5 days, IgG-containing cell supernatant from these cells was harvested by spinning twice at maximum speed ( > 4000 g) and subsequent filtration with 0.45 nm puradisc syringe filter (Whatmann, GE Healthcare, 10462100). .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Mass Spectrometry:

    Article Title: Poloxamer 188 decreases membrane toxicity of mutant SOD1 and ameliorates pathology observed in SOD1 mouse model for ALS
    Article Snippet: Proteins were desalted using 9K protein concentrators (Pierce, Rockford, IL) and stored in HBS (20 mM Hepes, 100 mM NaCl, pH 7.4) with 2.5% glycerol at −80° C. HBS and all solutions were made with BPC grade water (Sigma Aldrich, St. Louis, MO) unless stated otherwise. .. SOD1 metalation was measured by Perkin Elmer ELAN dynamic reactive cell inductively-coupled mass spectrometry performed using EPA-approved guidelines (method 1638; Brooks Rand Labs, Seattle).

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH). .. Recombinant VlpA7 (58.5 μg) was mixed with TiterMax Gold Adjuvant (Sigma Aldrich, St. Louis, MS) following the manufacturer's instructions, and injected subcutaneously into three C3H/HeNHsd mice (Harlan, Indianapolis, IN).

    Stable Transfection:

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: The full length plasmid containing α and β subunits was stably transfected in CHO cells via electroporation using the Gene Pulser Xcell system (BioRad). .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Synthesized:

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: VH and VL regions of PUMA1 were isolated by 5′ RACE, codon optimized and synthesized from Geneart (Life Technologies). .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    BIA-KA:

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: .. Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH). .. Recombinant VlpA7 (58.5 μg) was mixed with TiterMax Gold Adjuvant (Sigma Aldrich, St. Louis, MS) following the manufacturer's instructions, and injected subcutaneously into three C3H/HeNHsd mice (Harlan, Indianapolis, IN).

    Construct:

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: The constructs used for cell transfections were pPRGS827 (pCDNA3-SS::HA::MADD-4S::Gaussia Luciferase (GLc), obtained from P. Roy( )), pGM449 (pCMV::NLG-1::TM::FLAG::GFP), pGM451 (pCDNA3-SS::HA::NRX-1[EGF3+LNS6]), pGM467 (pCMV::NLG-1::His), pGM469 (pCDNA3-SS::HA::NRX-1[EGF3+LNS6]::TM::FLAG::GFP), pGM447 (pCMV::NLG-1::His::TM::GFP), pGM472 (pCMV::NLG-1::TM::GFP), pGM464 (pCDNA3-SS::HA::MADD-4S::FLAG). .. After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific).

    Enzyme-linked Immunosorbent Assay:

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: Soluble FcRn expression was assessed by anti-tag enzyme-linked immunosorbent assays (ELISA) and the 8 highest expressing CHO cell clones during early growth were pooled and scaled. .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Incubation:

    Article Title: In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import
    Article Snippet: Finally, bound antibodies were eluted twice by incubation for 10 min in 5 mL 0.1 M Glycine-HCl (pH 2.7), with eluates being immediately neutralized with 400 μL 2 M tris (pH 8.0) followed by dialysis in PBS overnight at 4°C using a 7K MWCO Pierce Slide-A-Lyzer® Dialysis Cassette (Thermo Fisher Scientific). .. Dialyzed eluates were combined and concentrated ~250-fold with a 9K MWCO Pierce Protein Concentrator (Thermo Fisher Scientific), and stored for later use at 4°C in 50% glycerol.

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: C. In log phase, cultures were induced with 0.35 mM IPTG and incubated at 30 deg. .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific).

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: In brief, a cell culture supernatant of FreeStyle 293-F cells transfected with the DARPin E2_79 expression vector pCH15 (PhCMV - SS SEAP -His-DARPin E2_79 -pA) was incubated overnight at 4 °C with equilibrated Ni-NTA agarose (cat. no. 745400.25; Macherey Nagel, Düren, Germany). .. Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS.

    Article Title: Inhibition of SIRT2 in merlin/NF2-mutant Schwann cells triggers necrosis
    Article Snippet: On the next day the medium was replaced with 2 ml treatment medium containing SIRT2 inhibitors/vehicle in duplicate wells and incubated for 6 hours at 37°C, 7% CO2. .. The supernatant (~4ml) was syringe filtered with 0.45 μm filter (Fisher), transferred to cold 7 ml 9K MWCO Pierce concentrators (Thermo Scientific) and centrifuged for 30 min at 4,000g, 4 °C.

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific). .. 48 hours posttransfection, culture media was removed and cells were incubated with concentrated conditioned media for 4 hours at 4°C.

    Amplification:

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: 2.9 Antibody generation The gene of the current Vmp expressed in our stock culture of B. hermsii DAH, vlpA7 , was amplified with P558 and P559, possessing NdeI and BamHI 5′ tails, respectively. .. Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH).

    Infection:

    Article Title: Novel Immunomodulators from Hard Ticks Selectively Reprogramme Human Dendritic Cell Responses
    Article Snippet: Protein purification The medium was cleared by centrifugation (2000 g, 10 min) 72 hours after infection, and proteins were precipitated by adding polyethylene glycol (PEG4000, Sigma; 18 g/100 ml). .. The protein-containing eluate fractions were pooled, concentrated using a 9K MWCO Pierce Protein Concentrator (Thermo Scientific), then further purified by size exclusion chromatography with a Superdex 75 HR10/30 column (GE Healthcare) using PBS (pH7.4) as running buffer.

    Expressing:

    Article Title: Phosphorylation of Synaptic GTPase-activating Protein (synGAP) by Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII) and Cyclin-dependent Kinase 5 (CDK5) Alters the Ratio of Its GAP Activity toward Ras and Rap GTPases *
    Article Snippet: Paragraph title: Cloning, Expression, and Purification of r-synGAP ... Fractions containing synGAP were pooled and concentrated by ultrafiltration through a 150-kDa cutoff filter (Thermo Scientific, catalog no. 89923) for r-synGAP, and a 9-kDa cutoff filter (Thermo Scientific, catalog no. 89885A) for sr-synGAP.

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: [ ] BL21(DE3) E. coli (New England Biolabs) were transformed with the expression plasmids and single colonies were grown in 1L 2XYT with 20% glucose at 37 deg. .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific).

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: In brief, a cell culture supernatant of FreeStyle 293-F cells transfected with the DARPin E2_79 expression vector pCH15 (PhCMV - SS SEAP -His-DARPin E2_79 -pA) was incubated overnight at 4 °C with equilibrated Ni-NTA agarose (cat. no. 745400.25; Macherey Nagel, Düren, Germany). .. Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS.

    Article Title: Poloxamer 188 decreases membrane toxicity of mutant SOD1 and ameliorates pathology observed in SOD1 mouse model for ALS
    Article Snippet: Paragraph title: Expression, purification and characterization of human SOD1 proteins ... Proteins were desalted using 9K protein concentrators (Pierce, Rockford, IL) and stored in HBS (20 mM Hepes, 100 mM NaCl, pH 7.4) with 2.5% glycerol at −80° C. HBS and all solutions were made with BPC grade water (Sigma Aldrich, St. Louis, MO) unless stated otherwise.

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: Soluble FcRn expression was assessed by anti-tag enzyme-linked immunosorbent assays (ELISA) and the 8 highest expressing CHO cell clones during early growth were pooled and scaled. .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: The variable region of the heavy chain (VH) was subcloned into pFuse expression vectors for mouse IgG1, IgG2a, IgG2b and IgG3 (Invivogen, pfuse-mg1fc1, pfuse-mg2afc1, pfuse-mg2bfc1, pfuse-mg3fc1); the variable region of the kappa light chain (VL) was subcloned into pFuse expression vector for mouse κ-Light chain (Invivogen, pfuse2-mclk). .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: To obtain concentrated conditioned culture media containing HA::MADD-4S::Gluc or NRX-1[EGF3+LNS6], 10 µg of each respective expression plasmid was transfected into HEK293T cells on 10-cm dishes, and culture media was changed to DMEM−0.2% FBS. .. After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific).

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The expression vector pGEX-6P-1, glutathione-Sepharose 4 Fast Flow, 3C protease (PreScission protease), HiLoad 16/600 Superdex 75 pg SEC column, and Mono Q 4.6/100 PE ion exchange column were obtained from GE Healthcare Life Sciences (Little Chalfont, UK). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

    Bradford Assay:

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS. .. The final protein concentration was determined using a Bradford assay (cat. no. 500-0006; Bio-Rad Laboratories, Inc., Hercules, CA, USA) and Coomassie blue gel staining.

    Modification:

    Article Title: Inhibition of SIRT2 in merlin/NF2-mutant Schwann cells triggers necrosis
    Article Snippet: The supernatant (~4ml) was syringe filtered with 0.45 μm filter (Fisher), transferred to cold 7 ml 9K MWCO Pierce concentrators (Thermo Scientific) and centrifuged for 30 min at 4,000g, 4 °C. .. When media was removed from cultures, the plates were washed once with ice cold PBS and cell were lysed with the modified RIPA buffer mentioned above; protein concentration was measured with the DC assay (BioRad).

    Western Blot:

    Article Title: Poloxamer 188 decreases membrane toxicity of mutant SOD1 and ameliorates pathology observed in SOD1 mouse model for ALS
    Article Snippet: Proteins were desalted using 9K protein concentrators (Pierce, Rockford, IL) and stored in HBS (20 mM Hepes, 100 mM NaCl, pH 7.4) with 2.5% glycerol at −80° C. HBS and all solutions were made with BPC grade water (Sigma Aldrich, St. Louis, MO) unless stated otherwise. .. Protein purity was determined by SDS-PAGE followed by Coomassie blue and silver staining, as well as by Western blots immunostained with antibody against WT- or mt-SOD1.

    Article Title: Inhibition of SIRT2 in merlin/NF2-mutant Schwann cells triggers necrosis
    Article Snippet: The supernatant (~4ml) was syringe filtered with 0.45 μm filter (Fisher), transferred to cold 7 ml 9K MWCO Pierce concentrators (Thermo Scientific) and centrifuged for 30 min at 4,000g, 4 °C. .. Equal volume of concentrated proteins from medium and 10 μg of protein of cell lysates were analyzed by western blotting for HMGB1.

    Article Title: The BH4 Domain of Anti-apoptotic Bcl-XL, but Not That of the Related Bcl-2, Limits the Voltage-dependent Anion Channel 1 (VDAC1)-mediated Transfer of Pro-apoptotic Ca2+ Signals to Mitochondria *
    Article Snippet: .. MAMs were centrifuged for 10 min at 7000 × g three times to remove mitochondrial contamination and finally centrifuged at 100000 × g for 1 h. The MAM fraction pellet was collected and concentrated using Pierce Protein Concentrators, 9K MWCO (Thermo Fisher Scientific Inc.) before Western blotting. .. For immunoprecipitation, 300 μg of protein lysate from COS-1 cells overexpressing Bcl-2 or Bcl-XL in the 3×FLAG pCMV-24 vector (Sigma) was added to 30 μl of anti-FLAG M2-agarose affinity gel (Sigma) previously washed according to the manufacturer's protocol.

    Transformation Assay:

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: [ ] BL21(DE3) E. coli (New England Biolabs) were transformed with the expression plasmids and single colonies were grown in 1L 2XYT with 20% glucose at 37 deg. .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific).

    Electroporation:

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: The full length plasmid containing α and β subunits was stably transfected in CHO cells via electroporation using the Gene Pulser Xcell system (BioRad). .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Transfection:

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: In brief, a cell culture supernatant of FreeStyle 293-F cells transfected with the DARPin E2_79 expression vector pCH15 (PhCMV - SS SEAP -His-DARPin E2_79 -pA) was incubated overnight at 4 °C with equilibrated Ni-NTA agarose (cat. no. 745400.25; Macherey Nagel, Düren, Germany). .. Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS.

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: The full length plasmid containing α and β subunits was stably transfected in CHO cells via electroporation using the Gene Pulser Xcell system (BioRad). .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: All IgGs were produced by transient transfection of HEK-freestyle cells (Thermo Fisher Scientific), as previously described by Vink et al. and Dekkers et al. After 5 days, IgG-containing cell supernatant from these cells was harvested by spinning twice at maximum speed ( > 4000 g) and subsequent filtration with 0.45 nm puradisc syringe filter (Whatmann, GE Healthcare, 10462100). .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: To obtain concentrated conditioned culture media containing HA::MADD-4S::Gluc or NRX-1[EGF3+LNS6], 10 µg of each respective expression plasmid was transfected into HEK293T cells on 10-cm dishes, and culture media was changed to DMEM−0.2% FBS. .. After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific).

    Gas Chromatography:

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: The constructs used for cell transfections were pPRGS827 (pCDNA3-SS::HA::MADD-4S::Gaussia Luciferase (GLc), obtained from P. Roy( )), pGM449 (pCMV::NLG-1::TM::FLAG::GFP), pGM451 (pCDNA3-SS::HA::NRX-1[EGF3+LNS6]), pGM467 (pCMV::NLG-1::His), pGM469 (pCDNA3-SS::HA::NRX-1[EGF3+LNS6]::TM::FLAG::GFP), pGM447 (pCMV::NLG-1::His::TM::GFP), pGM472 (pCMV::NLG-1::TM::GFP), pGM464 (pCDNA3-SS::HA::MADD-4S::FLAG). .. After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific).

    Protease Inhibitor:

    Article Title: Phosphorylation of Synaptic GTPase-activating Protein (synGAP) by Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII) and Cyclin-dependent Kinase 5 (CDK5) Alters the Ratio of Its GAP Activity toward Ras and Rap GTPases *
    Article Snippet: Cultures were then chilled to 18 °C, and protein expression was induced with 0.2 m m isopropyl 1-thio-β- d -galactopyranoside for 24 h. Bacterial pellets were harvested by centrifugation and resuspended in Lysis/Wash Buffer (20 m m Tris, 500 m m NaCl, 10 m m tris(2-carboxyethyl)phosphine (TCEP; Gold Biotechnology, catalog no. TCEP25), 5 m m MgCl2 , 60 m m imidazole, 2 m m ATP, 1 m m PMSF (Sigma, catalog no. ), 0.2% TergitolTM -type Nonidet P-40, and Complete EDTA-free protease inhibitor (Roche Applied Science, catalog no. 5056489001), pH 7.0) supplemented with 25 units/ml benzonase (EMD Millipore, catalog no. 71206) and 10 kilounits/ml ReadyLyse (Epicenter, catalog no. R1810M). .. Fractions containing synGAP were pooled and concentrated by ultrafiltration through a 150-kDa cutoff filter (Thermo Scientific, catalog no. 89923) for r-synGAP, and a 9-kDa cutoff filter (Thermo Scientific, catalog no. 89885A) for sr-synGAP.

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: Bacteria were pelleted, re-suspended in buffer containing 20 mM Tris, pH 7.4, 200 mM NaCl, 1 mM EDTA, 10 mM 2-mercaptoethanol and protease inhibitor cocktail (Sigma-Aldrich). .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific).

    Cell Culture:

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: In brief, a cell culture supernatant of FreeStyle 293-F cells transfected with the DARPin E2_79 expression vector pCH15 (PhCMV - SS SEAP -His-DARPin E2_79 -pA) was incubated overnight at 4 °C with equilibrated Ni-NTA agarose (cat. no. 745400.25; Macherey Nagel, Düren, Germany). .. Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS.

    Article Title: Inhibition of SIRT2 in merlin/NF2-mutant Schwann cells triggers necrosis
    Article Snippet: HMGB1 Release Assessment Equal number of merlin-null MSC per well in CellBIND 6-well plates were cultured overnight. .. The supernatant (~4ml) was syringe filtered with 0.45 μm filter (Fisher), transferred to cold 7 ml 9K MWCO Pierce concentrators (Thermo Scientific) and centrifuged for 30 min at 4,000g, 4 °C.

    SDS Page:

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific). .. All solid-phase resins, Fmoc-amino acids, and coupling reagents were purchased from EMD Biosciences unless otherwise noted.

    Article Title: Poloxamer 188 decreases membrane toxicity of mutant SOD1 and ameliorates pathology observed in SOD1 mouse model for ALS
    Article Snippet: Proteins were desalted using 9K protein concentrators (Pierce, Rockford, IL) and stored in HBS (20 mM Hepes, 100 mM NaCl, pH 7.4) with 2.5% glycerol at −80° C. HBS and all solutions were made with BPC grade water (Sigma Aldrich, St. Louis, MO) unless stated otherwise. .. Protein purity was determined by SDS-PAGE followed by Coomassie blue and silver staining, as well as by Western blots immunostained with antibody against WT- or mt-SOD1.

    Protein Concentration:

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS. .. The final protein concentration was determined using a Bradford assay (cat. no. 500-0006; Bio-Rad Laboratories, Inc., Hercules, CA, USA) and Coomassie blue gel staining.

    Article Title: Inhibition of SIRT2 in merlin/NF2-mutant Schwann cells triggers necrosis
    Article Snippet: The supernatant (~4ml) was syringe filtered with 0.45 μm filter (Fisher), transferred to cold 7 ml 9K MWCO Pierce concentrators (Thermo Scientific) and centrifuged for 30 min at 4,000g, 4 °C. .. When media was removed from cultures, the plates were washed once with ice cold PBS and cell were lysed with the modified RIPA buffer mentioned above; protein concentration was measured with the DC assay (BioRad).

    Polymerase Chain Reaction:

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The restriction endonucleases BamHI and XhoI, T4 DNA ligase, dNTP, GeneJET™ Gel Extraction Kit, GeneJET™ PCR Purification Kit, Phusion DNA Polymerase, and GeneJET™ Plasmid Miniprep Kit were acquired from Thermo Scientific (Waltham, MA, USA). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

    Sonication:

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: Cells were lysed by sonication and cell debris was pelleted. .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific).

    Injection:

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH). .. Recombinant VlpA7 (58.5 μg) was mixed with TiterMax Gold Adjuvant (Sigma Aldrich, St. Louis, MS) following the manufacturer's instructions, and injected subcutaneously into three C3H/HeNHsd mice (Harlan, Indianapolis, IN).

    Binding Assay:

    Article Title: In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import
    Article Snippet: Antibody purification using blotted recombinant GMD and GMER proteins To reduce nonspecific binding and cross-reactivity of GMD and GMER chicken IgY antibodies, 200 μg purified GMD/GMER protein was fractionated in 12.5% polyacrylamide gel and electroblotted for 1.5 h at 100 mA onto 0.2 μm nitrocellulose (Bio-Rad Laboratories) using a TE 77 Semi-Dry Transfer Unit (Hoefer, San Francisco, CA, USA). .. Dialyzed eluates were combined and concentrated ~250-fold with a 9K MWCO Pierce Protein Concentrator (Thermo Fisher Scientific), and stored for later use at 4°C in 50% glycerol.

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: Paragraph title: HEK293T cell surface binding experiments ... After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific).

    Molecular Weight:

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA). .. The protein molecular weight standards Precision Plus Protein Dual Xtra and Pageruler Unstained Low Range Protein Ladder were purchased from Bio-Rad (Hercules, CA, USA) and Thermo Scientific, respectively.

    Mutagenesis:

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The QuikChange Lightning Site-Directed Mutagenesis Kit was purchased from Stratagene (La Jolla, CA, USA). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

    Isolation:

    Article Title: In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import
    Article Snippet: This antibody isolation procedure was repeated twice more using the same antigen-bound strips. .. Dialyzed eluates were combined and concentrated ~250-fold with a 9K MWCO Pierce Protein Concentrator (Thermo Fisher Scientific), and stored for later use at 4°C in 50% glycerol.

    Article Title: Isolation and Characterization of the Herpes Simplex Virus 1 Terminase Complex
    Article Snippet: UL28 protein complexes were isolated from vFH476-infected Vero cells by a partial TAP procedure after elution from the streptavidin resin. .. The eluted protein sample was concentrated 10-fold by column centrifugation (Pierce; catalog no. 89884A).

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: IgG was isolated from cell supernatant with affinity chromatography columns HiTrap Protein A HP (GE Healthcare, 29–0485–76) for human IgG1, IgG2, and IgG4 or HiTrap Protein G HP (GE Healthcare, 29–0485–81) for human IgG3 and mouse IgG1, IgG2a, IgG2b and IgG3 on ÄKTA prime (GE Healthcare) according to standard procedures. .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Article Title: The BH4 Domain of Anti-apoptotic Bcl-XL, but Not That of the Related Bcl-2, Limits the Voltage-dependent Anion Channel 1 (VDAC1)-mediated Transfer of Pro-apoptotic Ca2+ Signals to Mitochondria *
    Article Snippet: Paragraph title: Isolation of MAMs ... MAMs were centrifuged for 10 min at 7000 × g three times to remove mitochondrial contamination and finally centrifuged at 100000 × g for 1 h. The MAM fraction pellet was collected and concentrated using Pierce Protein Concentrators, 9K MWCO (Thermo Fisher Scientific Inc.) before Western blotting.

    Silver Staining:

    Article Title: Poloxamer 188 decreases membrane toxicity of mutant SOD1 and ameliorates pathology observed in SOD1 mouse model for ALS
    Article Snippet: Proteins were desalted using 9K protein concentrators (Pierce, Rockford, IL) and stored in HBS (20 mM Hepes, 100 mM NaCl, pH 7.4) with 2.5% glycerol at −80° C. HBS and all solutions were made with BPC grade water (Sigma Aldrich, St. Louis, MO) unless stated otherwise. .. Protein purity was determined by SDS-PAGE followed by Coomassie blue and silver staining, as well as by Western blots immunostained with antibody against WT- or mt-SOD1.

    Flow Cytometry:

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: The heterodimeric, functional FcRn protein was purified from the media using IgG Sepharose 6 Fast Flow (GE Healthcare, 52–2083–00) on an Econo Column (BioRad, 737–2522). .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The expression vector pGEX-6P-1, glutathione-Sepharose 4 Fast Flow, 3C protease (PreScission protease), HiLoad 16/600 Superdex 75 pg SEC column, and Mono Q 4.6/100 PE ion exchange column were obtained from GE Healthcare Life Sciences (Little Chalfont, UK). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

    Mouse Assay:

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: Briefly, wild-type mice were transfused with KEL1 RBCs to induce immunity and a monoclonal anti-KEL1 antibody was isolated by standard myeloma fusion (PUMA1 antibody). .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH). .. Recombinant VlpA7 (58.5 μg) was mixed with TiterMax Gold Adjuvant (Sigma Aldrich, St. Louis, MS) following the manufacturer's instructions, and injected subcutaneously into three C3H/HeNHsd mice (Harlan, Indianapolis, IN).

    Protein Purification:

    Article Title: Novel Immunomodulators from Hard Ticks Selectively Reprogramme Human Dendritic Cell Responses
    Article Snippet: Paragraph title: Protein purification ... The protein-containing eluate fractions were pooled, concentrated using a 9K MWCO Pierce Protein Concentrator (Thermo Scientific), then further purified by size exclusion chromatography with a Superdex 75 HR10/30 column (GE Healthcare) using PBS (pH7.4) as running buffer.

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: The 1,016 base pair product was cloned into pET15b (Novagen‐Merck Millipore), and recombinant protein was obtained by Ni‐NTA agarose column protein purification (Qiagen). .. Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH).

    Recombinant:

    Article Title: In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import
    Article Snippet: Paragraph title: Antibody purification using blotted recombinant GMD and GMER proteins ... Dialyzed eluates were combined and concentrated ~250-fold with a 9K MWCO Pierce Protein Concentrator (Thermo Fisher Scientific), and stored for later use at 4°C in 50% glycerol.

    Article Title: Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence. Cis‐acting DNA elements flanking the variable major protein expression site of Borrelia hermsii are required for murine persistence
    Article Snippet: The 1,016 base pair product was cloned into pET15b (Novagen‐Merck Millipore), and recombinant protein was obtained by Ni‐NTA agarose column protein purification (Qiagen). .. Proteins were concentrated with Pierce Protein Concentrators 9K MWCO and quantified with a Micro BCA Protein Assay kit (Thermo Scientific, Marietta, OH).

    Staining:

    Article Title: In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import
    Article Snippet: Following transfer, the membrane-immobilized proteins were visualized with Ponceau S stain (Sigma-Aldrich), and bands were excised by razorblade. .. Dialyzed eluates were combined and concentrated ~250-fold with a 9K MWCO Pierce Protein Concentrator (Thermo Fisher Scientific), and stored for later use at 4°C in 50% glycerol.

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS. .. The final protein concentration was determined using a Bradford assay (cat. no. 500-0006; Bio-Rad Laboratories, Inc., Hercules, CA, USA) and Coomassie blue gel staining.

    Purification:

    Article Title: Novel Immunomodulators from Hard Ticks Selectively Reprogramme Human Dendritic Cell Responses
    Article Snippet: .. The protein-containing eluate fractions were pooled, concentrated using a 9K MWCO Pierce Protein Concentrator (Thermo Scientific), then further purified by size exclusion chromatography with a Superdex 75 HR10/30 column (GE Healthcare) using PBS (pH7.4) as running buffer. .. Concentration of purified protein was measured by its absorbance at 280 nm using extinction coefficients reported by the ProtParam tool ( http://web.expasy.org/protparam/ ).

    Article Title: Phosphorylation of Synaptic GTPase-activating Protein (synGAP) by Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII) and Cyclin-dependent Kinase 5 (CDK5) Alters the Ratio of Its GAP Activity toward Ras and Rap GTPases *
    Article Snippet: Paragraph title: Cloning, Expression, and Purification of r-synGAP ... Fractions containing synGAP were pooled and concentrated by ultrafiltration through a 150-kDa cutoff filter (Thermo Scientific, catalog no. 89923) for r-synGAP, and a 9-kDa cutoff filter (Thermo Scientific, catalog no. 89885A) for sr-synGAP.

    Article Title: In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import
    Article Snippet: Antibody purification using blotted recombinant GMD and GMER proteins To reduce nonspecific binding and cross-reactivity of GMD and GMER chicken IgY antibodies, 200 μg purified GMD/GMER protein was fractionated in 12.5% polyacrylamide gel and electroblotted for 1.5 h at 100 mA onto 0.2 μm nitrocellulose (Bio-Rad Laboratories) using a TE 77 Semi-Dry Transfer Unit (Hoefer, San Francisco, CA, USA). .. Dialyzed eluates were combined and concentrated ~250-fold with a 9K MWCO Pierce Protein Concentrator (Thermo Fisher Scientific), and stored for later use at 4°C in 50% glycerol.

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: Cleared lysate was diluted 1:2 in buffer and purified by amylose column chromatography (New England Biolabs). .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific).

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: Paragraph title: Purification of DARPin E2_79 ... Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS.

    Article Title: Poloxamer 188 decreases membrane toxicity of mutant SOD1 and ameliorates pathology observed in SOD1 mouse model for ALS
    Article Snippet: Paragraph title: Expression, purification and characterization of human SOD1 proteins ... Proteins were desalted using 9K protein concentrators (Pierce, Rockford, IL) and stored in HBS (20 mM Hepes, 100 mM NaCl, pH 7.4) with 2.5% glycerol at −80° C. HBS and all solutions were made with BPC grade water (Sigma Aldrich, St. Louis, MO) unless stated otherwise.

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots. .. Characterization of purified hFcRn FcRn protein purity was analyzed by SDS-Tris-glycine polyacrylamide gel electrophoresis (Life Technologies) under reducing conditions followed by SYPRO ruby staining (Life Technologies) using the manufacturers' recommended protocol.

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384). .. Antibody concentration was determined using Nanodrop 2000c UV/VIS spectrophotometer (Thermo Fisher Scientific).

    Article Title: The BH4 Domain of Anti-apoptotic Bcl-XL, but Not That of the Related Bcl-2, Limits the Voltage-dependent Anion Channel 1 (VDAC1)-mediated Transfer of Pro-apoptotic Ca2+ Signals to Mitochondria *
    Article Snippet: The homogenate was purified by centrifugation at 95,000 × g for 30 min on a PercollTM density gradient in isolation buffer 1 (a 15% Percoll gradient on a 30% layer). .. MAMs were centrifuged for 10 min at 7000 × g three times to remove mitochondrial contamination and finally centrifuged at 100000 × g for 1 h. The MAM fraction pellet was collected and concentrated using Pierce Protein Concentrators, 9K MWCO (Thermo Fisher Scientific Inc.) before Western blotting.

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The restriction endonucleases BamHI and XhoI, T4 DNA ligase, dNTP, GeneJET™ Gel Extraction Kit, GeneJET™ PCR Purification Kit, Phusion DNA Polymerase, and GeneJET™ Plasmid Miniprep Kit were acquired from Thermo Scientific (Waltham, MA, USA). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

    SPR Assay:

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384). .. Surface plasmon resonance (SPR) measurements were carried out on a IBIS MX96 (IBIS technologies) as described.

    Plasmid Preparation:

    Article Title: Sensing and responding to allergic response cytokines through a genetically encoded circuit
    Article Snippet: In brief, a cell culture supernatant of FreeStyle 293-F cells transfected with the DARPin E2_79 expression vector pCH15 (PhCMV - SS SEAP -His-DARPin E2_79 -pA) was incubated overnight at 4 °C with equilibrated Ni-NTA agarose (cat. no. 745400.25; Macherey Nagel, Düren, Germany). .. Finally, the DARPin E2_79 fraction was concentrated by ultrafiltration using concentrator columns (cat. no 89884A; Thermo Fisher Scientific, Waltham, MA, USA) and the elution buffer was successively replaced by PBS.

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: The full length plasmid containing α and β subunits was stably transfected in CHO cells via electroporation using the Gene Pulser Xcell system (BioRad). .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: The variable region of the heavy chain (VH) was subcloned into pFuse expression vectors for mouse IgG1, IgG2a, IgG2b and IgG3 (Invivogen, pfuse-mg1fc1, pfuse-mg2afc1, pfuse-mg2bfc1, pfuse-mg3fc1); the variable region of the kappa light chain (VL) was subcloned into pFuse expression vector for mouse κ-Light chain (Invivogen, pfuse2-mclk). .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Article Title: MADD-4/Punctin and Neurexin Organize C. elegans GABAergic Postsynapses through Neuroligin
    Article Snippet: To obtain concentrated conditioned culture media containing HA::MADD-4S::Gluc or NRX-1[EGF3+LNS6], 10 µg of each respective expression plasmid was transfected into HEK293T cells on 10-cm dishes, and culture media was changed to DMEM−0.2% FBS. .. After 24 hours of culture, the conditioned media was collected and centrifuged to remove cellular debris, and subsequently concentrated 10 times using Pierce Protein concentrators 9K MWCO (Thermo Scientific).

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The expression vector pGEX-6P-1, glutathione-Sepharose 4 Fast Flow, 3C protease (PreScission protease), HiLoad 16/600 Superdex 75 pg SEC column, and Mono Q 4.6/100 PE ion exchange column were obtained from GE Healthcare Life Sciences (Little Chalfont, UK). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

    Micro-arrays for Mass Spectrometry:

    Article Title: The BH4 Domain of Anti-apoptotic Bcl-XL, but Not That of the Related Bcl-2, Limits the Voltage-dependent Anion Channel 1 (VDAC1)-mediated Transfer of Pro-apoptotic Ca2+ Signals to Mitochondria *
    Article Snippet: .. MAMs were centrifuged for 10 min at 7000 × g three times to remove mitochondrial contamination and finally centrifuged at 100000 × g for 1 h. The MAM fraction pellet was collected and concentrated using Pierce Protein Concentrators, 9K MWCO (Thermo Fisher Scientific Inc.) before Western blotting. .. For immunoprecipitation, 300 μg of protein lysate from COS-1 cells overexpressing Bcl-2 or Bcl-XL in the 3×FLAG pCMV-24 vector (Sigma) was added to 30 μl of anti-FLAG M2-agarose affinity gel (Sigma) previously washed according to the manufacturer's protocol.

    Functional Assay:

    Article Title: A novel in vitro assay to predict neonatal Fc receptor-mediated human IgG half-life
    Article Snippet: The heterodimeric, functional FcRn protein was purified from the media using IgG Sepharose 6 Fast Flow (GE Healthcare, 52–2083–00) on an Econo Column (BioRad, 737–2522). .. Fractions were concentrated with centrifugal concentrators (Pierce, Thermo Scientific, 89885A) and purified FcRn was stored at −80°C as single-use aliquots.

    Affinity Chromatography:

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: IgG was isolated from cell supernatant with affinity chromatography columns HiTrap Protein A HP (GE Healthcare, 29–0485–76) for human IgG1, IgG2, and IgG4 or HiTrap Protein G HP (GE Healthcare, 29–0485–81) for human IgG3 and mouse IgG1, IgG2a, IgG2b and IgG3 on ÄKTA prime (GE Healthcare) according to standard procedures. .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Size-exclusion Chromatography:

    Article Title: Novel Immunomodulators from Hard Ticks Selectively Reprogramme Human Dendritic Cell Responses
    Article Snippet: .. The protein-containing eluate fractions were pooled, concentrated using a 9K MWCO Pierce Protein Concentrator (Thermo Scientific), then further purified by size exclusion chromatography with a Superdex 75 HR10/30 column (GE Healthcare) using PBS (pH7.4) as running buffer. .. Concentration of purified protein was measured by its absorbance at 280 nm using extinction coefficients reported by the ProtParam tool ( http://web.expasy.org/protparam/ ).

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The expression vector pGEX-6P-1, glutathione-Sepharose 4 Fast Flow, 3C protease (PreScission protease), HiLoad 16/600 Superdex 75 pg SEC column, and Mono Q 4.6/100 PE ion exchange column were obtained from GE Healthcare Life Sciences (Little Chalfont, UK). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

    Column Chromatography:

    Article Title: Peptide Bicycles that Inhibit the Grb2 SH2 Domain
    Article Snippet: Cleared lysate was diluted 1:2 in buffer and purified by amylose column chromatography (New England Biolabs). .. Fractions were analyzed by SDS-PAGE, pooled and concentrated using centrifugal 9K MWCO protein concentrators (Thermo Scientific).

    Antibody Purification:

    Article Title: In silico analysis of the fucosylation-associated genome of the human blood fluke Schistosoma mansoni: cloning and characterization of the enzymes involved in GDP-L-fucose synthesis and Golgi import
    Article Snippet: Paragraph title: Antibody purification using blotted recombinant GMD and GMER proteins ... Dialyzed eluates were combined and concentrated ~250-fold with a 9K MWCO Pierce Protein Concentrator (Thermo Fisher Scientific), and stored for later use at 4°C in 50% glycerol.

    Spectrophotometry:

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384). .. Antibody concentration was determined using Nanodrop 2000c UV/VIS spectrophotometer (Thermo Fisher Scientific).

    Produced:

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: All IgGs were produced by transient transfection of HEK-freestyle cells (Thermo Fisher Scientific), as previously described by Vink et al. and Dekkers et al. After 5 days, IgG-containing cell supernatant from these cells was harvested by spinning twice at maximum speed ( > 4000 g) and subsequent filtration with 0.45 nm puradisc syringe filter (Whatmann, GE Healthcare, 10462100). .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384).

    Concentration Assay:

    Article Title: Novel Immunomodulators from Hard Ticks Selectively Reprogramme Human Dendritic Cell Responses
    Article Snippet: The protein-containing eluate fractions were pooled, concentrated using a 9K MWCO Pierce Protein Concentrator (Thermo Scientific), then further purified by size exclusion chromatography with a Superdex 75 HR10/30 column (GE Healthcare) using PBS (pH7.4) as running buffer. .. Concentration of purified protein was measured by its absorbance at 280 nm using extinction coefficients reported by the ProtParam tool ( http://web.expasy.org/protparam/ ).

    Article Title: Affinity of human IgG subclasses to mouse Fc gamma receptors
    Article Snippet: .. Purified antibody fractions were concentrated to concentration > 1mg/mL using Protein Concentrators, 9K MWCO (Pierce, Thermo Fisher Scientific, 89884A) and subsequently dialysed against phosphate-buffered saline (PBS) overnight using Slide-A-Lyzer™ Dialysis Cassettes, 10K MWCO (Thermo Fisher Scientific, 66384). .. Antibody concentration was determined using Nanodrop 2000c UV/VIS spectrophotometer (Thermo Fisher Scientific).

    Lysis:

    Article Title: Phosphorylation of Synaptic GTPase-activating Protein (synGAP) by Ca2+/Calmodulin-dependent Protein Kinase II (CaMKII) and Cyclin-dependent Kinase 5 (CDK5) Alters the Ratio of Its GAP Activity toward Ras and Rap GTPases *
    Article Snippet: The resin was poured into a gravity-fed column and washed with 10 column volumes of Lysis/Wash Buffer. r-synGAP was eluted with Lysis/Wash Buffer (−2 m m ATP) supplemented with 10 m m EDTA and 250 m m imidazole. .. Fractions containing synGAP were pooled and concentrated by ultrafiltration through a 150-kDa cutoff filter (Thermo Scientific, catalog no. 89923) for r-synGAP, and a 9-kDa cutoff filter (Thermo Scientific, catalog no. 89885A) for sr-synGAP.

    Gel Extraction:

    Article Title: Mirolysin, a LysargiNase from Tannerella forsythia, proteolytically inactivates the human cathelicidin, LL-37
    Article Snippet: The restriction endonucleases BamHI and XhoI, T4 DNA ligase, dNTP, GeneJET™ Gel Extraction Kit, GeneJET™ PCR Purification Kit, Phusion DNA Polymerase, and GeneJET™ Plasmid Miniprep Kit were acquired from Thermo Scientific (Waltham, MA, USA). .. The Pierce Protein Concentrators 9K MWCO (7 ml and 20 ml) were from Thermo Scientific (Rockford, Il, USA).

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    Thermo Fisher pierce 9k mwco protein concentrators
    Pierce 9k Mwco Protein Concentrators, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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