liquid chromatography mass spectrometry  (Millipore)


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    Name:
    Liquid Chromatography
    Description:
    With the advent of new interfacing technology the benefits of LC MS liquid chromatography with mass spectrometry are being realized by the growth in applications of this technique in both the chemical and life sciences Topics covered include Limitations of the component techniques when used in isolation and how a combination of the two allows these to be overcome Descriptions of the various approaches including thermospray electrospray and atmospheric pressure chemical ionization which are used to interface the two techniques along with the advantages and disadvantages of each system Illustrative examples of the applications of LC MS including the molecular weight and structure determinations of both biopolymers and low molecular compounds
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    z702080
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    Structured Review

    Millipore liquid chromatography mass spectrometry
    Liquid Chromatography
    With the advent of new interfacing technology the benefits of LC MS liquid chromatography with mass spectrometry are being realized by the growth in applications of this technique in both the chemical and life sciences Topics covered include Limitations of the component techniques when used in isolation and how a combination of the two allows these to be overcome Descriptions of the various approaches including thermospray electrospray and atmospheric pressure chemical ionization which are used to interface the two techniques along with the advantages and disadvantages of each system Illustrative examples of the applications of LC MS including the molecular weight and structure determinations of both biopolymers and low molecular compounds
    https://www.bioz.com/result/liquid chromatography mass spectrometry/product/Millipore
    Average 95 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    liquid chromatography mass spectrometry - by Bioz Stars, 2020-05
    95/100 stars

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    Related Articles

    High Performance Liquid Chromatography:

    Article Title: Morusinol Exhibits Selective and Potent Antitumor Activity Against Human Liver Carcinoma by Inducing Autophagy, G2/M Cell Cycle Arrest, Inhibition of Cell Invasion and Migration, and Targeting of Ras/MEK/ERK Pathway
    Article Snippet: .. Chemicals and other reagents Morusinol (purity > 98%; determined by high-performance liquid chromatography), 3-(4, 5-dimethyl-2-thiazolyl)-2, and 5-diphenyl-2H-tetrazolium bromide (MTT) were obtained from SigmaAldrich Chemical Co. (St. Louis, MO, USA). .. Propidium iodide was purchased from Wuhan Boster Biological Technology (Wuhan, China).

    Article Title: Remodeling of Retinal Fatty Acids in an Animal Model of Diabetes
    Article Snippet: .. High-performance liquid chromatography (HPLC)-grade acetonitrile, acetic acid, methanol, chloroform, streptozotocin (STZ), and commonly used chemicals and reagents were from Sigma-Aldrich Chemical (St. Louis, MO). ..

    MTT Assay:

    Article Title: Morusinol Exhibits Selective and Potent Antitumor Activity Against Human Liver Carcinoma by Inducing Autophagy, G2/M Cell Cycle Arrest, Inhibition of Cell Invasion and Migration, and Targeting of Ras/MEK/ERK Pathway
    Article Snippet: .. Chemicals and other reagents Morusinol (purity > 98%; determined by high-performance liquid chromatography), 3-(4, 5-dimethyl-2-thiazolyl)-2, and 5-diphenyl-2H-tetrazolium bromide (MTT) were obtained from SigmaAldrich Chemical Co. (St. Louis, MO, USA). .. Propidium iodide was purchased from Wuhan Boster Biological Technology (Wuhan, China).

    Mass Spectrometry:

    Article Title: Enhanced dissolution and oral absorption of tacrolimus by supersaturable self-emulsifying drug delivery system
    Article Snippet: .. Ascomycin (purity > 98% w/w%) used as an internal standard for FK506 Liquid Chromatography-Tandem Mass Spectrometry (LC–MS/MS) analysis was obtained from Sigma-Aldrich Co. (St Louis, MO, USA). .. Cremophor EL and Soluplus were kindly provided by BASF (Ludwigshafen, Germany).

    Chromatography:

    Article Title: Enhanced dissolution and oral absorption of tacrolimus by supersaturable self-emulsifying drug delivery system
    Article Snippet: .. Ascomycin (purity > 98% w/w%) used as an internal standard for FK506 Liquid Chromatography-Tandem Mass Spectrometry (LC–MS/MS) analysis was obtained from Sigma-Aldrich Co. (St Louis, MO, USA). .. Cremophor EL and Soluplus were kindly provided by BASF (Ludwigshafen, Germany).

    Article Title: Formation of m2G6 in Methanocaldococcus jannaschii tRNA catalyzed by the novel methyltransferase Trm14
    Article Snippet: .. For liquid chromatography–mass spectrometry (LC–MS) analysis of modified nucleosides, nuclease P1 (Sigma), snake venom phosphodiesterase I (Worthington Biochemical Corporation) and Antarctic phosphatase (New England Biolabs) were used to digest Trm14-reacted transcripts and control unmethylated transcripts to nucleosides. .. The nucleosides were separated using a Hitachi D-7000 HPLC with a Hitachi L-7400 UV detector at 0.3 ml/min at room temperature on an LC-18 S 2.1 × 250 mm column from Supelco using a gradient of 5 mM ammonium acetate pH 5.3 and acetonitrile:water (40:60 v/v) as described ( ).

    Article Title: Unexpected Different Binding of Mistletoe Lectins from Plant Extracts to Immobilized Lactose and N-acetylgalactosamine
    Article Snippet: .. Fast protein liquid chromatography (FPLC) separation of eluted MLs using cation exchange chromatography The eluate from the affigel-asialofetuin column was dialysed against 15 mM citrate buffer (pH 4.2), then concentrated to 2 mg/ml using Microcon YM-30 tubes (Millipore, Switzerland) before applying to a cation exchange chromatography column (Mono S 5/50 GL; Amersham Bioscience, Switzerland). .. The column was adapted to a FPLC system and equilibrated with citrate buffer (pH 4.2).

    Isolation:

    Article Title: Attenuation of atherogenic apo B-48-dependent hyperlipidemia and high density lipoprotein remodeling induced by vitamin C and E combination and their beneficial effect on lethal ischemic heart disease in mice
    Article Snippet: .. Lipoprotein fractionation and high density lipoprotein isolation Serum was fractionated by fast-protein liquid chromatography (FPLC) and apolipoprotein A-I-containing fractions were pooled and concentrated using Amicon Ultra-4 centrifugal 10 K filters (Millipore, Merck, Darmstadt, HD, Germany). .. Pooled fractions were subjected to dynabead-based immunoprecipitation with an anti-apolipoprotein B (apo B) antibody to remove apo B-containing lipoproteins.

    Fractionation:

    Article Title: Attenuation of atherogenic apo B-48-dependent hyperlipidemia and high density lipoprotein remodeling induced by vitamin C and E combination and their beneficial effect on lethal ischemic heart disease in mice
    Article Snippet: .. Lipoprotein fractionation and high density lipoprotein isolation Serum was fractionated by fast-protein liquid chromatography (FPLC) and apolipoprotein A-I-containing fractions were pooled and concentrated using Amicon Ultra-4 centrifugal 10 K filters (Millipore, Merck, Darmstadt, HD, Germany). .. Pooled fractions were subjected to dynabead-based immunoprecipitation with an anti-apolipoprotein B (apo B) antibody to remove apo B-containing lipoproteins.

    Fast Protein Liquid Chromatography:

    Article Title: Attenuation of atherogenic apo B-48-dependent hyperlipidemia and high density lipoprotein remodeling induced by vitamin C and E combination and their beneficial effect on lethal ischemic heart disease in mice
    Article Snippet: .. Serum was fractionated by fast-protein liquid chromatography (FPLC) and apolipoprotein A-I-containing fractions were pooled and concentrated using Amicon Ultra-4 centrifugal 10 K filters (Millipore, Merck, Darmstadt, HD, Germany). .. Pooled fractions were subjected to dynabead-based immunoprecipitation with an anti-apolipoprotein B (apo B) antibody to remove apo B-containing lipoproteins.

    Article Title: Attenuation of atherogenic apo B-48-dependent hyperlipidemia and high density lipoprotein remodeling induced by vitamin C and E combination and their beneficial effect on lethal ischemic heart disease in mice
    Article Snippet: .. Lipoprotein fractionation and high density lipoprotein isolation Serum was fractionated by fast-protein liquid chromatography (FPLC) and apolipoprotein A-I-containing fractions were pooled and concentrated using Amicon Ultra-4 centrifugal 10 K filters (Millipore, Merck, Darmstadt, HD, Germany). .. Pooled fractions were subjected to dynabead-based immunoprecipitation with an anti-apolipoprotein B (apo B) antibody to remove apo B-containing lipoproteins.

    Article Title: Unexpected Different Binding of Mistletoe Lectins from Plant Extracts to Immobilized Lactose and N-acetylgalactosamine
    Article Snippet: .. Fast protein liquid chromatography (FPLC) separation of eluted MLs using cation exchange chromatography The eluate from the affigel-asialofetuin column was dialysed against 15 mM citrate buffer (pH 4.2), then concentrated to 2 mg/ml using Microcon YM-30 tubes (Millipore, Switzerland) before applying to a cation exchange chromatography column (Mono S 5/50 GL; Amersham Bioscience, Switzerland). .. The column was adapted to a FPLC system and equilibrated with citrate buffer (pH 4.2).

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: Formation of m2G6 in Methanocaldococcus jannaschii tRNA catalyzed by the novel methyltransferase Trm14
    Article Snippet: .. For liquid chromatography–mass spectrometry (LC–MS) analysis of modified nucleosides, nuclease P1 (Sigma), snake venom phosphodiesterase I (Worthington Biochemical Corporation) and Antarctic phosphatase (New England Biolabs) were used to digest Trm14-reacted transcripts and control unmethylated transcripts to nucleosides. .. The nucleosides were separated using a Hitachi D-7000 HPLC with a Hitachi L-7400 UV detector at 0.3 ml/min at room temperature on an LC-18 S 2.1 × 250 mm column from Supelco using a gradient of 5 mM ammonium acetate pH 5.3 and acetonitrile:water (40:60 v/v) as described ( ).

    Modification:

    Article Title: Formation of m2G6 in Methanocaldococcus jannaschii tRNA catalyzed by the novel methyltransferase Trm14
    Article Snippet: .. For liquid chromatography–mass spectrometry (LC–MS) analysis of modified nucleosides, nuclease P1 (Sigma), snake venom phosphodiesterase I (Worthington Biochemical Corporation) and Antarctic phosphatase (New England Biolabs) were used to digest Trm14-reacted transcripts and control unmethylated transcripts to nucleosides. .. The nucleosides were separated using a Hitachi D-7000 HPLC with a Hitachi L-7400 UV detector at 0.3 ml/min at room temperature on an LC-18 S 2.1 × 250 mm column from Supelco using a gradient of 5 mM ammonium acetate pH 5.3 and acetonitrile:water (40:60 v/v) as described ( ).

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  • 92
    Millipore maldi ms
    Characterization of ADP-ribosyl-HNP-1 from BAL fluid by <t>RP-HPLC,</t> <t>MALDI,</t> and enzymatic digestion. ( a ) Alignment of RP-HPLC chromatograms of products of ART-1-catalyzed ADP-ribosylation of HNP-1 (continuous line) and a chromatogram of a BAL sample from a smoker (dotted line). Arrows indicate elution times of ADP-ribosyl-HNP-1 and HNP-1. ( b ) ( i ) The peak with elution time compatible with ADP-ribosyl-HNP-1 (46.5 min), analyzed by MALDI-MS, shows a mass of 3,983 Da, consistent with ADP-ribosyl-HNP-1 (calculated 3,983 Da). ( ii ) Incubation of ADP-ribosyl-HNP-1 with pyrophosphatase and alkaline phosphatase produced ribosyl-HNP-1 (calculated 3,574 Da). ( iii ) ADP-ribosylarginine hydrolase cleaved the ribose-arginine linkage to release HNP-1 (calculated 3,443 Da). m / z is on the x axis.
    Maldi Ms, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/maldi ms/product/Millipore
    Average 92 stars, based on 3 article reviews
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    maldi ms - by Bioz Stars, 2020-05
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    92
    Millipore trastuzumab
    <t>Trastuzumab</t> and single hinge cleaved trastuzumab tumor inhibition efficacy in breast cancer xenograft mouse model . (A) BALB/c nu/nu mice ( n = 5) were subcutaneously inoculated with 5×10 6 BT474 human breast cancer cells. Mice were treated with the antibodies at 5 mg/kg weekly for a total of five doses when tumors reached an average size of 100 mm 3 . Tumor sizes were measured and compared among the treatment groups. * P
    Trastuzumab, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trastuzumab/product/Millipore
    Average 92 stars, based on 3 article reviews
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    86
    Millipore hpiv3 infected hela cell lysates
    Immunoprecipitation and flow cytometry assay. (A) Recombinant <t>GST-HPIV3-HN</t> or GST protein was immnoprecipitated with either #7, #21, #23 MAbs, or IgG (negative control), respectively. Then bound proteins were analyzed with immunoblotting using anti-GST antibody. (B) HPIV3-infected or uninfected <t>HeLa</t> cells were harvested at 4 days post-infection, followed by incubation with indicated MAbs. The cells were then fixed and stained with anti-mouse secondary antibody. The population of stained cells was calculated by flow cytometry. The shaded histogram shows negative hybridoma supernatant and the bold line shows specific MAbs.
    Hpiv3 Infected Hela Cell Lysates, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore amauroderma rude
    Identification of ergosterol A. The structure and composition of ergosterol analyzed by EI-MS. B. Structure of ergosterol based on the NMR data. C. Measurement of ergosterol concentrations from Ganoderma sinense, Ganoderma lucidum, <t>Amauroderma</t> rude, Coriolus versicolor , and Ganoderma tsugae .
    Amauroderma Rude, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of ADP-ribosyl-HNP-1 from BAL fluid by RP-HPLC, MALDI, and enzymatic digestion. ( a ) Alignment of RP-HPLC chromatograms of products of ART-1-catalyzed ADP-ribosylation of HNP-1 (continuous line) and a chromatogram of a BAL sample from a smoker (dotted line). Arrows indicate elution times of ADP-ribosyl-HNP-1 and HNP-1. ( b ) ( i ) The peak with elution time compatible with ADP-ribosyl-HNP-1 (46.5 min), analyzed by MALDI-MS, shows a mass of 3,983 Da, consistent with ADP-ribosyl-HNP-1 (calculated 3,983 Da). ( ii ) Incubation of ADP-ribosyl-HNP-1 with pyrophosphatase and alkaline phosphatase produced ribosyl-HNP-1 (calculated 3,574 Da). ( iii ) ADP-ribosylarginine hydrolase cleaved the ribose-arginine linkage to release HNP-1 (calculated 3,443 Da). m / z is on the x axis.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: ADP ribosylation of human neutrophil peptide-1 regulates its biological properties

    doi: 10.1073/pnas.122238899

    Figure Lengend Snippet: Characterization of ADP-ribosyl-HNP-1 from BAL fluid by RP-HPLC, MALDI, and enzymatic digestion. ( a ) Alignment of RP-HPLC chromatograms of products of ART-1-catalyzed ADP-ribosylation of HNP-1 (continuous line) and a chromatogram of a BAL sample from a smoker (dotted line). Arrows indicate elution times of ADP-ribosyl-HNP-1 and HNP-1. ( b ) ( i ) The peak with elution time compatible with ADP-ribosyl-HNP-1 (46.5 min), analyzed by MALDI-MS, shows a mass of 3,983 Da, consistent with ADP-ribosyl-HNP-1 (calculated 3,983 Da). ( ii ) Incubation of ADP-ribosyl-HNP-1 with pyrophosphatase and alkaline phosphatase produced ribosyl-HNP-1 (calculated 3,574 Da). ( iii ) ADP-ribosylarginine hydrolase cleaved the ribose-arginine linkage to release HNP-1 (calculated 3,443 Da). m / z is on the x axis.

    Article Snippet: We incubated ADP-ribosyl-HNP-1 (6 pmol/0.2 μg) in 200 μl of 250 mM NaHCO3 /25 mM MgCl2 , alkaline phosphatase (5 μg), and pyrophosphatase (5 μg) (Sigma) for 30 min at 37°C before termination of the reaction with 6 M guanidine, separation of reaction products by RP-HPLC, and analysis by MALDI-MS. We also incubated ADP-ribosyl-HNP-1 (11 pmol, 0.42 μg) overnight at 37°C with recombinant human ADP-ribosylarginine hydrolase (1 μg) in 100 μl of 50 mM Tris (pH 7.5)/5 mM DTT/10 mM Mg Cl2 , followed by desalting and concentration using Zip-TipC18 (Millipore), and analysis by MALDI-MS.

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry, Incubation, Produced

    Trastuzumab and single hinge cleaved trastuzumab tumor inhibition efficacy in breast cancer xenograft mouse model . (A) BALB/c nu/nu mice ( n = 5) were subcutaneously inoculated with 5×10 6 BT474 human breast cancer cells. Mice were treated with the antibodies at 5 mg/kg weekly for a total of five doses when tumors reached an average size of 100 mm 3 . Tumor sizes were measured and compared among the treatment groups. * P

    Journal: Breast Cancer Research : BCR

    Article Title: A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy

    doi: 10.1186/bcr3240

    Figure Lengend Snippet: Trastuzumab and single hinge cleaved trastuzumab tumor inhibition efficacy in breast cancer xenograft mouse model . (A) BALB/c nu/nu mice ( n = 5) were subcutaneously inoculated with 5×10 6 BT474 human breast cancer cells. Mice were treated with the antibodies at 5 mg/kg weekly for a total of five doses when tumors reached an average size of 100 mm 3 . Tumor sizes were measured and compared among the treatment groups. * P

    Article Snippet: Flow cytometry Binding of trastuzumab and scIgG-T on HER2-expressing breast cancer cells was measured using a Guava easyCyte HT instrument based on the manufacturer's instructions (Millipore, Danvers, MA, USA).

    Techniques: Inhibition, Mouse Assay

    Detection of single hinge cleaved trastuzumab and matrix metalloproteinases in breast cancer patient samples . (A) Same amounts of enriched IgGs from each sample were separated by SDS-PAGE under nonreducing/denaturing conditions. Arrow, single hinge cleaved trastuzumab (scIgG-T) as referenced to standard preparation in the right lane. (B) Detection of matrix metalloproteinase (MMP) expression in breast cancer patient tumor tissues and normal adjacent 2 cm tissue using a reverse protein array method ( n = 4). * P

    Journal: Breast Cancer Research : BCR

    Article Title: A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy

    doi: 10.1186/bcr3240

    Figure Lengend Snippet: Detection of single hinge cleaved trastuzumab and matrix metalloproteinases in breast cancer patient samples . (A) Same amounts of enriched IgGs from each sample were separated by SDS-PAGE under nonreducing/denaturing conditions. Arrow, single hinge cleaved trastuzumab (scIgG-T) as referenced to standard preparation in the right lane. (B) Detection of matrix metalloproteinase (MMP) expression in breast cancer patient tumor tissues and normal adjacent 2 cm tissue using a reverse protein array method ( n = 4). * P

    Article Snippet: Flow cytometry Binding of trastuzumab and scIgG-T on HER2-expressing breast cancer cells was measured using a Guava easyCyte HT instrument based on the manufacturer's instructions (Millipore, Danvers, MA, USA).

    Techniques: SDS Page, Expressing, Protein Array

    Detection of single hinge cleaved trastuzumab and trastuzumab proteolytic cleavage using mass spectrometry . (A) Sequence of the hinge region of trastuzumab showing the major trypsin cleavage sites (black arrows) and the IgG-degrading enzyme S (IdeS) cleavage site (red arrow). Bottom graph: liquid chromatography (LC)/mass spectrometry (MS)/MS analysis of trypsin-treated single hinge cleaved trastuzumab (scIgG-T), identifying the two peptides: THTCPPCPAPELLG and GPSVFLFPPKPK. Spectra are the MS2 spectra showing the b and y ions for the two peptides, confirming their sequences. (B) Sequence of the hinge region of trastuzumab. Black arrows, major trypsin cleavage sites. Bottom graph: LC/MS/MS analysis of trypsin-treated trastuzumab that identifies the peptide THTCPPCPAPELLGGPSVFLFPPKPK. Spectra are the MS2 spectra showing the b and y ions for the peptide, confirming its sequence.

    Journal: Breast Cancer Research : BCR

    Article Title: A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy

    doi: 10.1186/bcr3240

    Figure Lengend Snippet: Detection of single hinge cleaved trastuzumab and trastuzumab proteolytic cleavage using mass spectrometry . (A) Sequence of the hinge region of trastuzumab showing the major trypsin cleavage sites (black arrows) and the IgG-degrading enzyme S (IdeS) cleavage site (red arrow). Bottom graph: liquid chromatography (LC)/mass spectrometry (MS)/MS analysis of trypsin-treated single hinge cleaved trastuzumab (scIgG-T), identifying the two peptides: THTCPPCPAPELLG and GPSVFLFPPKPK. Spectra are the MS2 spectra showing the b and y ions for the two peptides, confirming their sequences. (B) Sequence of the hinge region of trastuzumab. Black arrows, major trypsin cleavage sites. Bottom graph: LC/MS/MS analysis of trypsin-treated trastuzumab that identifies the peptide THTCPPCPAPELLGGPSVFLFPPKPK. Spectra are the MS2 spectra showing the b and y ions for the peptide, confirming its sequence.

    Article Snippet: Flow cytometry Binding of trastuzumab and scIgG-T on HER2-expressing breast cancer cells was measured using a Guava easyCyte HT instrument based on the manufacturer's instructions (Millipore, Danvers, MA, USA).

    Techniques: Mass Spectrometry, Sequencing, Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy

    Biological characterization of single hinge cleaved trastuzumab compared with the intact trastuzumab antibody in vitro . (A) Histograms of trastuzumab and single hinge cleaved trastuzumab (scIgG-T) binding to HER2 expressed on BT474 cells using a flow cytometer. (B) Concentration-dependent binding of trastuzumab and scIgG-T to HER2 expressed on BT474 breast cancer cells as measured by flow cytometer. Mean fluorescence intensity (MFI) is plotted against each antibody concentration (nM) on the x axis. (C) Effect of trastuzumab and scIgG-T on total HER2 expression, pHER2 (Y1289), pAKT (S473), and pErk1/2 in BT474 cells as determined by western blotting. (D) Inhibition of BT474 breast cancer cell proliferation by trastuzumab and scIgG-T ( n = 4). Percentage of cell growth inhibition calculated as: (fluorescence signal of control group - signal of treatment group)/signal of control group×100.

    Journal: Breast Cancer Research : BCR

    Article Title: A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy

    doi: 10.1186/bcr3240

    Figure Lengend Snippet: Biological characterization of single hinge cleaved trastuzumab compared with the intact trastuzumab antibody in vitro . (A) Histograms of trastuzumab and single hinge cleaved trastuzumab (scIgG-T) binding to HER2 expressed on BT474 cells using a flow cytometer. (B) Concentration-dependent binding of trastuzumab and scIgG-T to HER2 expressed on BT474 breast cancer cells as measured by flow cytometer. Mean fluorescence intensity (MFI) is plotted against each antibody concentration (nM) on the x axis. (C) Effect of trastuzumab and scIgG-T on total HER2 expression, pHER2 (Y1289), pAKT (S473), and pErk1/2 in BT474 cells as determined by western blotting. (D) Inhibition of BT474 breast cancer cell proliferation by trastuzumab and scIgG-T ( n = 4). Percentage of cell growth inhibition calculated as: (fluorescence signal of control group - signal of treatment group)/signal of control group×100.

    Article Snippet: Flow cytometry Binding of trastuzumab and scIgG-T on HER2-expressing breast cancer cells was measured using a Guava easyCyte HT instrument based on the manufacturer's instructions (Millipore, Danvers, MA, USA).

    Techniques: In Vitro, Binding Assay, Flow Cytometry, Cytometry, Concentration Assay, Fluorescence, Expressing, Western Blot, Inhibition

    Trastuzumab and single hinge cleaved trastuzumab binding to Fcγ receptors and antibody-dependent cellular cytotoxicity activity . (A) Trastuzumab and single hinge cleaved trastuzumab (scIgG-T) binding to Fc gamma receptor (FcγR) I. (B) Trastuzumab and scIgG-T binding to FcγRIIA. (C) Trastuzumab and scIgG-T binding to FcγRIIIA. (D) Antibody-dependent cellular cytotoxicity (ADCC) activity induced by trastuzumab and scIgG-T using the high-HER2-expressing SKOV-3 ovarian cancer cells as target cells and human peripheral blood mononuclear cells (PBMC) as effector cells ( n = 3). Cell index was compared after treatment of trastuzumab and scIgG-T for 24 hours. Percentage of cell lysis calculated as: (cell index of control group - cell index of treatment group)/cell index of control group×100. RFU, relative fluorescence units.

    Journal: Breast Cancer Research : BCR

    Article Title: A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy

    doi: 10.1186/bcr3240

    Figure Lengend Snippet: Trastuzumab and single hinge cleaved trastuzumab binding to Fcγ receptors and antibody-dependent cellular cytotoxicity activity . (A) Trastuzumab and single hinge cleaved trastuzumab (scIgG-T) binding to Fc gamma receptor (FcγR) I. (B) Trastuzumab and scIgG-T binding to FcγRIIA. (C) Trastuzumab and scIgG-T binding to FcγRIIIA. (D) Antibody-dependent cellular cytotoxicity (ADCC) activity induced by trastuzumab and scIgG-T using the high-HER2-expressing SKOV-3 ovarian cancer cells as target cells and human peripheral blood mononuclear cells (PBMC) as effector cells ( n = 3). Cell index was compared after treatment of trastuzumab and scIgG-T for 24 hours. Percentage of cell lysis calculated as: (cell index of control group - cell index of treatment group)/cell index of control group×100. RFU, relative fluorescence units.

    Article Snippet: Flow cytometry Binding of trastuzumab and scIgG-T on HER2-expressing breast cancer cells was measured using a Guava easyCyte HT instrument based on the manufacturer's instructions (Millipore, Danvers, MA, USA).

    Techniques: Binding Assay, Activity Assay, Expressing, Lysis, Fluorescence

    Immune cell infiltration in xenograft tumors measured by immunohistochemistry . Mouse xenograft tumors treated with trastuzumab, single hinge cleaved trastuzumab (scIgG-T), and the isotype control were fixed in 4% paraformaldehyde. Six immunohistochemistry slides were prepared from the paraffin-embedded tissues of two mice from each treatment group. Monocytes/macrophages were stained using the anti-mouse integrin αM/CD11b and anti-mouse F4/80 antibodies. (A) Representative images (×40) of CD11b-stained tumor tissue and positive-stained immune cells indicated by black arrows. Right bar graph: average number of CD11b-positive cells per tumor tissue slide ( n = 6). Pairwise t test used to compare different treatment groups. ** P

    Journal: Breast Cancer Research : BCR

    Article Title: A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy

    doi: 10.1186/bcr3240

    Figure Lengend Snippet: Immune cell infiltration in xenograft tumors measured by immunohistochemistry . Mouse xenograft tumors treated with trastuzumab, single hinge cleaved trastuzumab (scIgG-T), and the isotype control were fixed in 4% paraformaldehyde. Six immunohistochemistry slides were prepared from the paraffin-embedded tissues of two mice from each treatment group. Monocytes/macrophages were stained using the anti-mouse integrin αM/CD11b and anti-mouse F4/80 antibodies. (A) Representative images (×40) of CD11b-stained tumor tissue and positive-stained immune cells indicated by black arrows. Right bar graph: average number of CD11b-positive cells per tumor tissue slide ( n = 6). Pairwise t test used to compare different treatment groups. ** P

    Article Snippet: Flow cytometry Binding of trastuzumab and scIgG-T on HER2-expressing breast cancer cells was measured using a Guava easyCyte HT instrument based on the manufacturer's instructions (Millipore, Danvers, MA, USA).

    Techniques: Immunohistochemistry, Mouse Assay, Staining

    Detection of trastuzumab proteolytic cleavage in vitro . (A) Capillary electropherograms (CEs) of trastuzumab and matrix metalloproteinase (MMP)-digested trastuzumab under nonreducing and denatured running conditions. (B) CE of trastuzumab and IgG-degrading enzyme S (IdeS)-digested single hinge cleaved trastuzumab (scIgG-T). (C) Purified scIgG-T under nonreducing and denaturing running conditions and stained with Coomassie blue. (D) Trastuzumab and scIgG-T under reducing and denaturing running conditions and stained with Coomassie blue. Fab, fragment antigen binding; Fc (m) , Fc monomer; HC, heavy chain; LC, light chain.

    Journal: Breast Cancer Research : BCR

    Article Title: A single proteolytic cleavage within the lower hinge of trastuzumab reduces immune effector function and in vivo efficacy

    doi: 10.1186/bcr3240

    Figure Lengend Snippet: Detection of trastuzumab proteolytic cleavage in vitro . (A) Capillary electropherograms (CEs) of trastuzumab and matrix metalloproteinase (MMP)-digested trastuzumab under nonreducing and denatured running conditions. (B) CE of trastuzumab and IgG-degrading enzyme S (IdeS)-digested single hinge cleaved trastuzumab (scIgG-T). (C) Purified scIgG-T under nonreducing and denaturing running conditions and stained with Coomassie blue. (D) Trastuzumab and scIgG-T under reducing and denaturing running conditions and stained with Coomassie blue. Fab, fragment antigen binding; Fc (m) , Fc monomer; HC, heavy chain; LC, light chain.

    Article Snippet: Flow cytometry Binding of trastuzumab and scIgG-T on HER2-expressing breast cancer cells was measured using a Guava easyCyte HT instrument based on the manufacturer's instructions (Millipore, Danvers, MA, USA).

    Techniques: In Vitro, Purification, Staining, Binding Assay

    Immunoprecipitation and flow cytometry assay. (A) Recombinant GST-HPIV3-HN or GST protein was immnoprecipitated with either #7, #21, #23 MAbs, or IgG (negative control), respectively. Then bound proteins were analyzed with immunoblotting using anti-GST antibody. (B) HPIV3-infected or uninfected HeLa cells were harvested at 4 days post-infection, followed by incubation with indicated MAbs. The cells were then fixed and stained with anti-mouse secondary antibody. The population of stained cells was calculated by flow cytometry. The shaded histogram shows negative hybridoma supernatant and the bold line shows specific MAbs.

    Journal: Frontiers in Microbiology

    Article Title: Wheat germ cell-free system-based production of hemagglutinin-neuraminidase glycoprotein of human parainfluenza virus type 3 for generation and characterization of monoclonal antibody

    doi: 10.3389/fmicb.2014.00208

    Figure Lengend Snippet: Immunoprecipitation and flow cytometry assay. (A) Recombinant GST-HPIV3-HN or GST protein was immnoprecipitated with either #7, #21, #23 MAbs, or IgG (negative control), respectively. Then bound proteins were analyzed with immunoblotting using anti-GST antibody. (B) HPIV3-infected or uninfected HeLa cells were harvested at 4 days post-infection, followed by incubation with indicated MAbs. The cells were then fixed and stained with anti-mouse secondary antibody. The population of stained cells was calculated by flow cytometry. The shaded histogram shows negative hybridoma supernatant and the bold line shows specific MAbs.

    Article Snippet: IMMUNOBLOTTING Recombinant HPIV3-HN proteins (equivalent to ~100 ng) or HPIV3-infected HeLa cell lysates were separated by 10% SDS-Gel and transferred onto a PVDF membrane (Millipore).

    Techniques: Immunoprecipitation, Flow Cytometry, Cytometry, Recombinant, Negative Control, Infection, Incubation, Staining

    Immunoblotting and immunofluorescent analysis. (A,B) Detection sensitivity of the MAbs for recombinant His-HN (A) or HPIV3-infected cell lysate (B) . Recombinant HPIV3-HN (100 ng) was separated using 12.5% SDS-gel and transferred to a PVDF membrane, followed by incubation with MAbs (hybridoma supernatants) at a 1:10 dilution (A) . HeLa cells were infected or mock-infected with HPIV3. After 48 h, cells were lysed with SDS-PAGE loading buffer. The total protein was separated in 12.5% SDS-gel and immunobloted with indicated MAbs (B) . (C,D) Immunofluoresent analysis of HN (red) in HPIV3-infected HeLa cells. HeLa cells were infected or mock-infected with HPIV3. After 48 h, cells were fixed, and then stained with MAbs (hybridoma supernatant; red) and DAPI (blue). Confocal microscopic analysis was performed at 40× (C) and at 600× magnifications (D) .

    Journal: Frontiers in Microbiology

    Article Title: Wheat germ cell-free system-based production of hemagglutinin-neuraminidase glycoprotein of human parainfluenza virus type 3 for generation and characterization of monoclonal antibody

    doi: 10.3389/fmicb.2014.00208

    Figure Lengend Snippet: Immunoblotting and immunofluorescent analysis. (A,B) Detection sensitivity of the MAbs for recombinant His-HN (A) or HPIV3-infected cell lysate (B) . Recombinant HPIV3-HN (100 ng) was separated using 12.5% SDS-gel and transferred to a PVDF membrane, followed by incubation with MAbs (hybridoma supernatants) at a 1:10 dilution (A) . HeLa cells were infected or mock-infected with HPIV3. After 48 h, cells were lysed with SDS-PAGE loading buffer. The total protein was separated in 12.5% SDS-gel and immunobloted with indicated MAbs (B) . (C,D) Immunofluoresent analysis of HN (red) in HPIV3-infected HeLa cells. HeLa cells were infected or mock-infected with HPIV3. After 48 h, cells were fixed, and then stained with MAbs (hybridoma supernatant; red) and DAPI (blue). Confocal microscopic analysis was performed at 40× (C) and at 600× magnifications (D) .

    Article Snippet: IMMUNOBLOTTING Recombinant HPIV3-HN proteins (equivalent to ~100 ng) or HPIV3-infected HeLa cell lysates were separated by 10% SDS-Gel and transferred onto a PVDF membrane (Millipore).

    Techniques: Recombinant, Infection, SDS-Gel, Incubation, SDS Page, Staining

    Proteomics analysis using selected MAb. (A) Schematic diagram of proteomic analysis for the identification of PIV3-HN-binding protein. HPIV3-infected HeLa cell lysate was immunoprecipitated with #21 MAb. The bound proteins were separated by SDS-PAGE and analyzed by LC-Ms/Ms. (B) The panel shows the list of putative HN-binding proteins identified by mass spectrometry analysis. (C) FLAG-tagged HSP70, HSP90, tubulin alpha 1C, or SERPINA3 proteins were mixed with HPIV3-HN. Samples were pull-down with the streptavidin magnetic beads and the collected proteins were separated by SDS-PAGE. The bound protein detected by immunoblotting analysis with anti-FLAG antibody. The right arrows indicated the position of each protein.

    Journal: Frontiers in Microbiology

    Article Title: Wheat germ cell-free system-based production of hemagglutinin-neuraminidase glycoprotein of human parainfluenza virus type 3 for generation and characterization of monoclonal antibody

    doi: 10.3389/fmicb.2014.00208

    Figure Lengend Snippet: Proteomics analysis using selected MAb. (A) Schematic diagram of proteomic analysis for the identification of PIV3-HN-binding protein. HPIV3-infected HeLa cell lysate was immunoprecipitated with #21 MAb. The bound proteins were separated by SDS-PAGE and analyzed by LC-Ms/Ms. (B) The panel shows the list of putative HN-binding proteins identified by mass spectrometry analysis. (C) FLAG-tagged HSP70, HSP90, tubulin alpha 1C, or SERPINA3 proteins were mixed with HPIV3-HN. Samples were pull-down with the streptavidin magnetic beads and the collected proteins were separated by SDS-PAGE. The bound protein detected by immunoblotting analysis with anti-FLAG antibody. The right arrows indicated the position of each protein.

    Article Snippet: IMMUNOBLOTTING Recombinant HPIV3-HN proteins (equivalent to ~100 ng) or HPIV3-infected HeLa cell lysates were separated by 10% SDS-Gel and transferred onto a PVDF membrane (Millipore).

    Techniques: Binding Assay, Infection, Immunoprecipitation, SDS Page, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Magnetic Beads

    Identification of ergosterol A. The structure and composition of ergosterol analyzed by EI-MS. B. Structure of ergosterol based on the NMR data. C. Measurement of ergosterol concentrations from Ganoderma sinense, Ganoderma lucidum, Amauroderma rude, Coriolus versicolor , and Ganoderma tsugae .

    Journal: Oncotarget

    Article Title: Ergosterol purified from medicinal mushroom Amauroderma rude inhibits cancer growth in vitro and in vivo by up-regulating multiple tumor suppressors

    doi:

    Figure Lengend Snippet: Identification of ergosterol A. The structure and composition of ergosterol analyzed by EI-MS. B. Structure of ergosterol based on the NMR data. C. Measurement of ergosterol concentrations from Ganoderma sinense, Ganoderma lucidum, Amauroderma rude, Coriolus versicolor , and Ganoderma tsugae .

    Article Snippet: Western blotting After being treated with the purified compound of Amauroderma rude , cells were lysed by the lysis buffer containing protease inhibitor (CALBIOCHEM, USA) on ice for 30 min and centrifuged to obtain clear cell lysates.

    Techniques: Mass Spectrometry, Nuclear Magnetic Resonance

    Purification of anti-cancer ingredient and molecule from Amauroderma rude The dry fruit bodies of Amauroderma rude were extracted using 95% ethanol. The extract was subject to chromatography and HPLC purification as indicated. Human breast cancer cell cultures were used to screen the anti-cancer ingredients in each step until single pick was obtained from HPLC. AReth, ethanol extract of Amauroderma rude ; ARchl, chloroform fraction of Amauroderma rude ; P/A, ratios of petroleum ether to ether in the elution buffer; Meth, pure methanol used to elute the column; Fr, fraction number; TL, fractions obtained from the Thin layer Chromatography; inset, photo of a dry Amauroderma rude .

    Journal: Oncotarget

    Article Title: Ergosterol purified from medicinal mushroom Amauroderma rude inhibits cancer growth in vitro and in vivo by up-regulating multiple tumor suppressors

    doi:

    Figure Lengend Snippet: Purification of anti-cancer ingredient and molecule from Amauroderma rude The dry fruit bodies of Amauroderma rude were extracted using 95% ethanol. The extract was subject to chromatography and HPLC purification as indicated. Human breast cancer cell cultures were used to screen the anti-cancer ingredients in each step until single pick was obtained from HPLC. AReth, ethanol extract of Amauroderma rude ; ARchl, chloroform fraction of Amauroderma rude ; P/A, ratios of petroleum ether to ether in the elution buffer; Meth, pure methanol used to elute the column; Fr, fraction number; TL, fractions obtained from the Thin layer Chromatography; inset, photo of a dry Amauroderma rude .

    Article Snippet: Western blotting After being treated with the purified compound of Amauroderma rude , cells were lysed by the lysis buffer containing protease inhibitor (CALBIOCHEM, USA) on ice for 30 min and centrifuged to obtain clear cell lysates.

    Techniques: Purification, Chromatography, High Performance Liquid Chromatography, Thin Layer Chromatography