protease inhibitor  (Millipore)

 
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    Name:
    Protease Inhibitor Mix
    Description:
    Protease inhibitor mix
    Catalog Number:
    GE80-6501-23
    Price:
    None
    Applications:
    Sample preparation often requires the inhibition of protease activity. GE Healthcare offers this combination of competitive and noncompetitive protease inhibitors, which protect Proteins from proteolysis during purification from animal tissues, plant tissues, yeast and bacteria.
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    Structured Review

    Millipore protease inhibitor
    Protease Inhibitor Mix
    Protease inhibitor mix
    https://www.bioz.com/result/protease inhibitor/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor - by Bioz Stars, 2021-05
    99/100 stars

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    Related Articles

    Recombinant:

    Article Title: P2X7 Receptor Activation Impairs Exogenous MHC Class I Oligopeptides Presentation in Antigen Presenting Cells
    Article Snippet: Escherichia coli LPS serotype 055:B5, ATP, MG132, etoposide, apyrase, pan-caspase inhibitor (Q-VD-OPh) and Brefeldin A were purchased from Sigma-Aldrich. .. Recombinant mouse IL-4 was from BD; caspase-3 inhibitor I (Ac-DEVD-CHO), GM6001 and E64 protease inhibitor from Calbiochem. .. P2X7R antagonists A438079 or A740003, the ecto-NTPDase inhibitor sodium polyoxotungstate (POM1), the ecto-5′-nucleotidase inhibitor adenosine 5′-(α,β-ethylene)diphosphate (APCP) and N ,N ,N ',N '-Tetrakis (2-pyridylmethyl)ethylenediamine (TPEN) from Tocris.

    Protease Inhibitor:

    Article Title: P2X7 Receptor Activation Impairs Exogenous MHC Class I Oligopeptides Presentation in Antigen Presenting Cells
    Article Snippet: Escherichia coli LPS serotype 055:B5, ATP, MG132, etoposide, apyrase, pan-caspase inhibitor (Q-VD-OPh) and Brefeldin A were purchased from Sigma-Aldrich. .. Recombinant mouse IL-4 was from BD; caspase-3 inhibitor I (Ac-DEVD-CHO), GM6001 and E64 protease inhibitor from Calbiochem. .. P2X7R antagonists A438079 or A740003, the ecto-NTPDase inhibitor sodium polyoxotungstate (POM1), the ecto-5′-nucleotidase inhibitor adenosine 5′-(α,β-ethylene)diphosphate (APCP) and N ,N ,N ',N '-Tetrakis (2-pyridylmethyl)ethylenediamine (TPEN) from Tocris.

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: .. Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). ..

    Article Title: Autophagic Inhibition via Lysosomal Integrity Dysfunction Leads to Antitumor Activity in Glioma Treatment
    Article Snippet: .. Bafilomycin A1, rapamycin, dithiothreitol (DTT), trifluoperazine, iodoacetamide (IAA), formic acid, E64D cysteine protease inhibitor, pifithrin-μ (PES), and artesunate (ART) were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. Pronase and protease inhibitor cocktail tablets were obtained from Roche (Basel, Switzerland).

    Article Title: Eps15 Homology Domain-containing Protein 3 Regulates Cardiac T-type Ca2+ Channel Targeting and Function in the Atria *
    Article Snippet: .. Protein A-conjugated agarose beads (AffiGel; Bio-Rad) were incubated with either control mouse Ig or anti-EHD3 Ig in co-immunoprecipitation binding buffer (PBS with 0.1% Triton X-100 and protease inhibitor mixture (Sigma)) for 12 h at 4 °C. ..

    Article Title: Sex-dependent role of microglia in disulfide high mobility group box 1 protein-mediated mechanical hypersensitivity
    Article Snippet: Drugs and drug deliveryEndotoxin-free, disulfide HMGB1 was kindly provided by Dr. H. Yang (Feinstein Institute for Medical Research, NY) or purchased from HMGBiotech (Milan, Italy). .. Minocycline (glial inhibitor), alpha-1-antitrypsin (A1AT, protease inhibitor), haptoglobin (sequesters HMGB1) and sivelestat (neutrophil elastase inhibitor) were all purchased from Sigma-Aldrich (St. Louis, MO). ..

    Article Title: Dianhydrogalactitol synergizes with topoisomerase poisons to overcome DNA repair activity in tumor cells
    Article Snippet: Tail Moments are presented as mean ± standard deviation. .. Western blot analysisCells were lysed with EBC buffer (50 mM Tris-HCl, pH 8.0, 120 mM NaCl, 1% NP-40, and 1 mM ethylenediaminetetraacetic acid; EDTA) supplemented with PhosSTOP phosphatase inhibitor and cOmplete protease inhibitor (MilliporeSigma, Oakville, Canada). ..

    Article Title: Strategy and rationale for urine collection protocols employed in the NEPTUNE study
    Article Snippet: FIBRO was well preserved in exosomes stored with or without protease inhibitor, whereas it was not well detected in cell pellets under these conditions, especially with storage. .. Scramblase (PLSCR1) was well preserved with Sigma protease inhibitor and one freeze/thaw cycle. .. As with cellular pellets, in general, addition of protease inhibitor was essential to protein survival (Fig. , Additional file : Figure S1).

    Article Title: Strategy and rationale for urine collection protocols employed in the NEPTUNE study
    Article Snippet: Cellular pellet proteins We also examined protein stability in cellular pellets for selected proteins using Western analysis, with and without the above protease inhibitors (Additional file : Figure S1). .. There was substantially better detection of the podocyte marker of interest PODXL in cellular pellets stored with protease inhibitors, and again found that Sigma protease inhibitor product may in fact be superior to the more expensive Roche protease inhibitor cocktail (Additional file : Figure S1). .. Without the presence of protease inhibitor, the proteins we examined (podocalyxin, fibrocystin and smoothened) did not survive 12 months of storage (6 month representative Western shown), or a freeze/thaw cycle at −80 °C.

    Affinity Purification:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: .. Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). ..

    Incubation:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: .. Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). ..

    Article Title: Eps15 Homology Domain-containing Protein 3 Regulates Cardiac T-type Ca2+ Channel Targeting and Function in the Atria *
    Article Snippet: .. Protein A-conjugated agarose beads (AffiGel; Bio-Rad) were incubated with either control mouse Ig or anti-EHD3 Ig in co-immunoprecipitation binding buffer (PBS with 0.1% Triton X-100 and protease inhibitor mixture (Sigma)) for 12 h at 4 °C. ..

    Binding Assay:

    Article Title: Eps15 Homology Domain-containing Protein 3 Regulates Cardiac T-type Ca2+ Channel Targeting and Function in the Atria *
    Article Snippet: .. Protein A-conjugated agarose beads (AffiGel; Bio-Rad) were incubated with either control mouse Ig or anti-EHD3 Ig in co-immunoprecipitation binding buffer (PBS with 0.1% Triton X-100 and protease inhibitor mixture (Sigma)) for 12 h at 4 °C. ..

    Western Blot:

    Article Title: Dianhydrogalactitol synergizes with topoisomerase poisons to overcome DNA repair activity in tumor cells
    Article Snippet: Tail Moments are presented as mean ± standard deviation. .. Western blot analysisCells were lysed with EBC buffer (50 mM Tris-HCl, pH 8.0, 120 mM NaCl, 1% NP-40, and 1 mM ethylenediaminetetraacetic acid; EDTA) supplemented with PhosSTOP phosphatase inhibitor and cOmplete protease inhibitor (MilliporeSigma, Oakville, Canada). ..

    Marker:

    Article Title: Strategy and rationale for urine collection protocols employed in the NEPTUNE study
    Article Snippet: Cellular pellet proteins We also examined protein stability in cellular pellets for selected proteins using Western analysis, with and without the above protease inhibitors (Additional file : Figure S1). .. There was substantially better detection of the podocyte marker of interest PODXL in cellular pellets stored with protease inhibitors, and again found that Sigma protease inhibitor product may in fact be superior to the more expensive Roche protease inhibitor cocktail (Additional file : Figure S1). .. Without the presence of protease inhibitor, the proteins we examined (podocalyxin, fibrocystin and smoothened) did not survive 12 months of storage (6 month representative Western shown), or a freeze/thaw cycle at −80 °C.

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  • 99
    Millipore western blot analysis transfected cells
    Associations with F are altered by O-glycan loss. A) Cell lysates of cells <t>transfected</t> with wt F and either wt or mutant HeV G. HeV G (top) is blotted with rabbit anti-HA, and HeV F (bottom) is blotted with mouse anti-AU1. B) Pulldown of HeV G and co-IP of F for transfected cells. Antibody blotting was performed as in A). C) Cell lysates of cells transfected with wt F and wt or mutant NiV G. D) Co-IP of NiV transfected cells. E) Co-IP values of HeV and NiV O-glycan mutants, determined by densitometry. Image Lab software (Biorad) was used to measure the densitometry of Western blot bands. Co-IP values were determined by dividing either the F 0 , F 1 , or total F densitometry values for the co-IP bands by the co-IP G densitometry values to account for any differences in co-IP G pulldown. These values were then divided by F 0 , F 1 , or total F densitometry values for cell lysates to account for differences in F mutant expression. Values were then normalized to those of wt HNV G. Averages are shown. N = 3. Statistically significant differences, as determined by a Student’s t-test, are marked with an * (p
    Western Blot Analysis Transfected Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/western blot analysis transfected cells/product/Millipore
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    95
    Millipore streptavidin horseradish peroxide hrp
    (A) Production of SpeB by A8173 and NZ131 wild-type bacteria and the NZ131 speB and NZ131 rgg SpeB-defective mutants. Bacteria from overnight cultures were lysed in SDS-PAGE sample buffer, and the samples subjected to SDS-PAGE and transferred to a nitrocellulose membrane, and then the membrane was probed with SpeB antiserum. (B) Strepadhesin activity of wild-type and SpeB-defective mutant streptococci. A membrane containing 1-μl aliquots of serially diluted (OD 600 = 4.0 to 0.125) suspensions of A8173 and NZ131 wild-type bacteria and the SpeB-defective NZ131 speB and NZ131 rgg mutants was incubated with biotinylated thyroglobulin, and the binding was detected with <t>streptavidin-HRP</t> and by ECL autoradiography.
    Streptavidin Horseradish Peroxide Hrp, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin horseradish peroxide hrp/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    93
    Millipore rat heart cytochrome c
    Representative immunoblots of left ventricular tissue from 2 sham, 2 ventricular fibrillation (VF) 4-min, and 2 VF 8-min groups showing <t>cytochrome</t> c and prohibitin in cytosolic and mitochondrial fractions and β-actin in cytosolic fraction. The corresponding densitometries for the entire group are shown in graphs depicting cytochrome c in mitochondrial fraction indexed to prohibitin ( left ) and cytochrome c in cytosolic fraction indexed to β-actin ( right ). White columns denote sham ( n = 8), gray columns denote VF 4-min ( n = 8), and black columns denote VF 8-min ( n = 8). NC, negative control. Means are ± SE. * P
    Rat Heart Cytochrome C, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat heart cytochrome c/product/Millipore
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    99
    Millipore recombinant lipin 1 proteins
    <t>Lipin-1</t> as a protein phosphatase-1c interaction partner through a potential RV X F-like motif. A , sequence alignment and secondary structure prediction of the N termini of Mus musculus lipins and the Saccharomyces cerevisiae Pah1p. Cylinders and arrows
    Recombinant Lipin 1 Proteins, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant lipin 1 proteins/product/Millipore
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    Image Search Results


    Associations with F are altered by O-glycan loss. A) Cell lysates of cells transfected with wt F and either wt or mutant HeV G. HeV G (top) is blotted with rabbit anti-HA, and HeV F (bottom) is blotted with mouse anti-AU1. B) Pulldown of HeV G and co-IP of F for transfected cells. Antibody blotting was performed as in A). C) Cell lysates of cells transfected with wt F and wt or mutant NiV G. D) Co-IP of NiV transfected cells. E) Co-IP values of HeV and NiV O-glycan mutants, determined by densitometry. Image Lab software (Biorad) was used to measure the densitometry of Western blot bands. Co-IP values were determined by dividing either the F 0 , F 1 , or total F densitometry values for the co-IP bands by the co-IP G densitometry values to account for any differences in co-IP G pulldown. These values were then divided by F 0 , F 1 , or total F densitometry values for cell lysates to account for differences in F mutant expression. Values were then normalized to those of wt HNV G. Averages are shown. N = 3. Statistically significant differences, as determined by a Student’s t-test, are marked with an * (p

    Journal: PLoS Pathogens

    Article Title: Multiple Novel Functions of Henipavirus O-glycans: The First O-glycan Functions Identified in the Paramyxovirus Family

    doi: 10.1371/journal.ppat.1005445

    Figure Lengend Snippet: Associations with F are altered by O-glycan loss. A) Cell lysates of cells transfected with wt F and either wt or mutant HeV G. HeV G (top) is blotted with rabbit anti-HA, and HeV F (bottom) is blotted with mouse anti-AU1. B) Pulldown of HeV G and co-IP of F for transfected cells. Antibody blotting was performed as in A). C) Cell lysates of cells transfected with wt F and wt or mutant NiV G. D) Co-IP of NiV transfected cells. E) Co-IP values of HeV and NiV O-glycan mutants, determined by densitometry. Image Lab software (Biorad) was used to measure the densitometry of Western blot bands. Co-IP values were determined by dividing either the F 0 , F 1 , or total F densitometry values for the co-IP bands by the co-IP G densitometry values to account for any differences in co-IP G pulldown. These values were then divided by F 0 , F 1 , or total F densitometry values for cell lysates to account for differences in F mutant expression. Values were then normalized to those of wt HNV G. Averages are shown. N = 3. Statistically significant differences, as determined by a Student’s t-test, are marked with an * (p

    Article Snippet: Western blot analysis Transfected cells or pseudotyped virions expressing HNV G wt or mutants with or without HNV F were lysed in RIPA buffer (Millipore) supplemented with complete protease inhibitor (cOmplete Mini, Roche).

    Techniques: Transfection, Mutagenesis, Co-Immunoprecipitation Assay, Software, Western Blot, Expressing

    (A) Production of SpeB by A8173 and NZ131 wild-type bacteria and the NZ131 speB and NZ131 rgg SpeB-defective mutants. Bacteria from overnight cultures were lysed in SDS-PAGE sample buffer, and the samples subjected to SDS-PAGE and transferred to a nitrocellulose membrane, and then the membrane was probed with SpeB antiserum. (B) Strepadhesin activity of wild-type and SpeB-defective mutant streptococci. A membrane containing 1-μl aliquots of serially diluted (OD 600 = 4.0 to 0.125) suspensions of A8173 and NZ131 wild-type bacteria and the SpeB-defective NZ131 speB and NZ131 rgg mutants was incubated with biotinylated thyroglobulin, and the binding was detected with streptavidin-HRP and by ECL autoradiography.

    Journal: Infection and Immunity

    Article Title: Streptococcus pyogenes Glycoprotein-Binding Strepadhesin Activity Is Mediated by a Surface-Associated Carbohydrate-Degrading Enzyme, Pullulanase

    doi: 10.1128/IAI.71.2.784-793.2003

    Figure Lengend Snippet: (A) Production of SpeB by A8173 and NZ131 wild-type bacteria and the NZ131 speB and NZ131 rgg SpeB-defective mutants. Bacteria from overnight cultures were lysed in SDS-PAGE sample buffer, and the samples subjected to SDS-PAGE and transferred to a nitrocellulose membrane, and then the membrane was probed with SpeB antiserum. (B) Strepadhesin activity of wild-type and SpeB-defective mutant streptococci. A membrane containing 1-μl aliquots of serially diluted (OD 600 = 4.0 to 0.125) suspensions of A8173 and NZ131 wild-type bacteria and the SpeB-defective NZ131 speB and NZ131 rgg mutants was incubated with biotinylated thyroglobulin, and the binding was detected with streptavidin-HRP and by ECL autoradiography.

    Article Snippet: Bovine thyroglobulin, fetuin, asialofetuin, and submaxillary mucin, horse myoglobin, streptavidin-horseradish peroxide (HRP), the cysteine protease inhibitor trans-epoxysuccinyl -l- leucylamido-(4-guanidino)butane (E64), and human placenta laminin were purchased from Sigma.

    Techniques: SDS Page, Activity Assay, Mutagenesis, Incubation, Binding Assay, Autoradiography

    Representative immunoblots of left ventricular tissue from 2 sham, 2 ventricular fibrillation (VF) 4-min, and 2 VF 8-min groups showing cytochrome c and prohibitin in cytosolic and mitochondrial fractions and β-actin in cytosolic fraction. The corresponding densitometries for the entire group are shown in graphs depicting cytochrome c in mitochondrial fraction indexed to prohibitin ( left ) and cytochrome c in cytosolic fraction indexed to β-actin ( right ). White columns denote sham ( n = 8), gray columns denote VF 4-min ( n = 8), and black columns denote VF 8-min ( n = 8). NC, negative control. Means are ± SE. * P

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Activation of caspase-3 may not contribute to postresuscitation myocardial dysfunction

    doi: 10.1152/ajpheart.00338.2008

    Figure Lengend Snippet: Representative immunoblots of left ventricular tissue from 2 sham, 2 ventricular fibrillation (VF) 4-min, and 2 VF 8-min groups showing cytochrome c and prohibitin in cytosolic and mitochondrial fractions and β-actin in cytosolic fraction. The corresponding densitometries for the entire group are shown in graphs depicting cytochrome c in mitochondrial fraction indexed to prohibitin ( left ) and cytochrome c in cytosolic fraction indexed to β-actin ( right ). White columns denote sham ( n = 8), gray columns denote VF 4-min ( n = 8), and black columns denote VF 8-min ( n = 8). NC, negative control. Means are ± SE. * P

    Article Snippet: CaCl2 , 3-[(3-cholamidopropyl)dimethyl ammonio]-1-propane sulfonate (CHAPS), DTT, EDTA, EGTA, glucose, HEPES, KCl, mannitol, MOPS, NaCl, paraformaldehyde, PMSF, proteinase K, protease inhibitor cocktail, phosphatase inhibitor cocktail, rat heart cytochrome c , RNase A (DNase free), SDS, NaF, Na3 VO4 , sucrose, Triton X-100, and triphenyltetrazolium chloride (TTC) were purchased from Sigma; N -acetyl-Asp-Glu-Val-Asp-7-amino-4-trifluoromethyl coumarin (Ac-DEVD-AFC; caspase-3 substrate) and DEVD-CHO (reversible caspase-3 inhibitor) were from Biomol; 1-mm-thick 12% and 14% Novex tris-glycine polyacrylamide gels were from Invitrogen; complete protease inhibitor cocktail tablets and polyvinylidene difluoride (PVDF) membrane were from Roche Applied Science; West femto maximum sensitivity chemiluminescent detection kit was from Pierce Biotechnology; protein concentration assay BCA kit based on the Bradford method ( ) was from Bio-Rad; APO-DNA1 PCR kit for DNA ladder assay was from Maxim Biotech; T4 -DNA ligase was from New England Biolabs; phase lock gel tubes were from Eppendorf; and bromophenol blue, ethidium bromide, Evan's blue, glycerol, isopropanol, phenol/chloroform/isoamyl alcohol, 2-mercaptoethanol, and Tris were from Fisher Biotech.

    Techniques: Western Blot, Negative Control

    Lipin-1 as a protein phosphatase-1c interaction partner through a potential RV X F-like motif. A , sequence alignment and secondary structure prediction of the N termini of Mus musculus lipins and the Saccharomyces cerevisiae Pah1p. Cylinders and arrows

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: Lipin-1 as a protein phosphatase-1c interaction partner through a potential RV X F-like motif. A , sequence alignment and secondary structure prediction of the N termini of Mus musculus lipins and the Saccharomyces cerevisiae Pah1p. Cylinders and arrows

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Sequencing

    Preincubation with PP-1cγ-interacting peptides prevents the association of lipin-1 with PP-1cγ. A , the amino acid sequences of synthetic peptides known to compete against PP-1c regulatory subunits (ZAP WT peptide) as well as a short peptide

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: Preincubation with PP-1cγ-interacting peptides prevents the association of lipin-1 with PP-1cγ. A , the amino acid sequences of synthetic peptides known to compete against PP-1c regulatory subunits (ZAP WT peptide) as well as a short peptide

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques:

    UV-circular dichroism spectra of lipin-1 wild type and HARA mutant. UV-circular dichroism analysis was performed on the recombinant purified FLAG-tagged lipin-1 wild type and the HARA mutant.

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: UV-circular dichroism spectra of lipin-1 wild type and HARA mutant. UV-circular dichroism analysis was performed on the recombinant purified FLAG-tagged lipin-1 wild type and the HARA mutant.

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Mutagenesis, Recombinant, Purification

    The effects of different lipin-1 mutations on its interaction with PP-1cγ. A , HEK 293 cell lysates expressing HA-tagged lipin-1 wild type, HA-lipin-1 phosphomimetic mutant (21S/T to E), and HA-lipin-1 truncation mutant (residues 321–2775)

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: The effects of different lipin-1 mutations on its interaction with PP-1cγ. A , HEK 293 cell lysates expressing HA-tagged lipin-1 wild type, HA-lipin-1 phosphomimetic mutant (21S/T to E), and HA-lipin-1 truncation mutant (residues 321–2775)

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Expressing, Mutagenesis

    Protein dot blots and Western blot of HEK 293 cell lysates overexpressing lipin-1 proteins. A , representative protein dot blots using 0.1–1 μg of protein from the HEK 293 cell lysates. The results show the expression of different FLAG-tagged

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: Protein dot blots and Western blot of HEK 293 cell lysates overexpressing lipin-1 proteins. A , representative protein dot blots using 0.1–1 μg of protein from the HEK 293 cell lysates. The results show the expression of different FLAG-tagged

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Western Blot, Expressing

    The effects of other lipin-1 NLIP point mutations on subcellular localization. A , confocal images of HEK 293 cells overexpressing lipin-1 21S/T to A or several representative 21S/T to A NLIP point mutants. B , quantification of the proportion of cells

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: The effects of other lipin-1 NLIP point mutations on subcellular localization. A , confocal images of HEK 293 cells overexpressing lipin-1 21S/T to A or several representative 21S/T to A NLIP point mutants. B , quantification of the proportion of cells

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques:

    Venn diagram depicting the effect of the different mutations in the lipin-1 N terminus on the interplay between PAP activity, the ability to interact with PP-1c, and nuclear localization. Our results demonstrate that the lipin-1 wild type and non-phosphorylatable

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: Venn diagram depicting the effect of the different mutations in the lipin-1 N terminus on the interplay between PAP activity, the ability to interact with PP-1c, and nuclear localization. Our results demonstrate that the lipin-1 wild type and non-phosphorylatable

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Activity Assay

    PAP activity of recombinant lipin-1 proteins. PAP activities of HEK 293 cell lysates overexpressing equivalent amounts of recombinant lipin-1 proteins are shown for three independent experiments. Error bars , S.E.

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: PAP activity of recombinant lipin-1 proteins. PAP activities of HEK 293 cell lysates overexpressing equivalent amounts of recombinant lipin-1 proteins are shown for three independent experiments. Error bars , S.E.

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Activity Assay, Recombinant

    Effect of HARA mutation on lipin-1 subcellular localization is not dependent on its effects on catalytic activity. A , confocal images showing the subcellular localization of recombinant lipin-1 wild type, HARA, D712E,D714E catalytically inactive mutant,

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: Effect of HARA mutation on lipin-1 subcellular localization is not dependent on its effects on catalytic activity. A , confocal images showing the subcellular localization of recombinant lipin-1 wild type, HARA, D712E,D714E catalytically inactive mutant,

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Mutagenesis, Activity Assay, Recombinant

    Lipin-1 binds to protein phosphatase-1. A , human embryonic kidney 293 (HEK 293) cell lysate overexpressing FLAG-tagged recombinant lipin-1 wild type was incubated with recombinant PP-1cγ bound to microcystin-LR-linked Sepharose beads ( MC-LR ;

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Residues in the N Terminus of Lipin-1 Are Required for Binding to Protein Phosphatase-1c, Nuclear Translocation, and Phosphatidate Phosphatase Activity *

    doi: 10.1074/jbc.M114.552612

    Figure Lengend Snippet: Lipin-1 binds to protein phosphatase-1. A , human embryonic kidney 293 (HEK 293) cell lysate overexpressing FLAG-tagged recombinant lipin-1 wild type was incubated with recombinant PP-1cγ bound to microcystin-LR-linked Sepharose beads ( MC-LR ;

    Article Snippet: HEK 293 cells overexpressing recombinant lipin-1 proteins were sonicated in 25 m m HEPES, pH 7.4, containing 250 m m sucrose, 2 m m DTT, protease inhibitor mixture (Sigma-Aldrich), 1 m m MnCl2 , 30 n m microcystin-LR, and 0.1% (w/v) Tween 20.

    Techniques: Recombinant, Incubation