dithiothreitol dtt  (Millipore)


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    Structured Review

    Millipore dithiothreitol dtt
    Dithiothreitol Dtt, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dithiothreitol dtt/product/Millipore
    Average 99 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    dithiothreitol dtt - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    High Performance Liquid Chromatography:

    Article Title: Protein Identification Pipeline for the Homology Driven Proteomics
    Article Snippet: Cleland’s reagent (DTT) and iodoacetamide were of analytical grade and purchased from Sigma-Aldrich (Munich, Germany); water and acetonitrile were of LC-MS grade from Fisher Scientific (Schwerte, Germany). .. Formic acid and trifluoroacetic acid were HPLC grade obtained from Merck (Darmstadt, Germany).

    Article Title: Oxidized Low-Density Lipoprotein Is a Novel Predictor of Interferon Responsiveness in Chronic Hepatitis C Infection
    Article Snippet: Linearized DNA was purified by phenol/chloroform extraction (cat. no. 0038.1, Roti-Phenol: Carl Roth, Karlsruhe, Germany; chloroform, cat. no. 3313.1: Carl Roth), precipitated with ethanol, and dissolved in high-performance liquid chromatography (HPCL) water (cat. no. A511.2; Carl Roth). .. The in vitro transcription reaction mixture contained 80 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, pH 7.5] (cat. no. HN77.1; Carl Roth) 12 mM MgCl2 (cat. no. HNO3.1; Carl Roth), 2 mM spermidine (cat. no. SO266-5G; Sigma-Aldrich), 40 mM dithiothreitol (DTT) (cat. no. 43915-5G; Sigma-Aldrich), a 3.125 mM concentration of each nucleoside triphosphate (cat. no. 11140957001, 11140922001, 11140965001, 11140949001; Roche, Basel, Switzerland), 1 U of RNasin/mL (cat. no. N2515; Promega, Mannheim, Germany), 0.1 μg of plasmid DNA/μL, and 0.6 U of T7 RNA polymerase/μL (cat. no. P2077; Promega).

    In Vitro:

    Article Title: Oxidized Low-Density Lipoprotein Is a Novel Predictor of Interferon Responsiveness in Chronic Hepatitis C Infection
    Article Snippet: .. The in vitro transcription reaction mixture contained 80 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, pH 7.5] (cat. no. HN77.1; Carl Roth) 12 mM MgCl2 (cat. no. HNO3.1; Carl Roth), 2 mM spermidine (cat. no. SO266-5G; Sigma-Aldrich), 40 mM dithiothreitol (DTT) (cat. no. 43915-5G; Sigma-Aldrich), a 3.125 mM concentration of each nucleoside triphosphate (cat. no. 11140957001, 11140922001, 11140965001, 11140949001; Roche, Basel, Switzerland), 1 U of RNasin/mL (cat. no. N2515; Promega, Mannheim, Germany), 0.1 μg of plasmid DNA/μL, and 0.6 U of T7 RNA polymerase/μL (cat. no. P2077; Promega). .. After 2 hours of incubation at 37°C, 0.3 U T7 RNA polymerase was added to the mixture and incubated for another 2 hours at 37°C.

    Modification:

    Article Title: Protein Identification Pipeline for the Homology Driven Proteomics
    Article Snippet: Cleland’s reagent (DTT) and iodoacetamide were of analytical grade and purchased from Sigma-Aldrich (Munich, Germany); water and acetonitrile were of LC-MS grade from Fisher Scientific (Schwerte, Germany). .. Modified porcine trypsin (sequencing grade) was purchased from Promega (Mannheim, Germany).

    Purification:

    Article Title: Oxidized Low-Density Lipoprotein Is a Novel Predictor of Interferon Responsiveness in Chronic Hepatitis C Infection
    Article Snippet: Linearized DNA was purified by phenol/chloroform extraction (cat. no. 0038.1, Roti-Phenol: Carl Roth, Karlsruhe, Germany; chloroform, cat. no. 3313.1: Carl Roth), precipitated with ethanol, and dissolved in high-performance liquid chromatography (HPCL) water (cat. no. A511.2; Carl Roth). .. The in vitro transcription reaction mixture contained 80 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, pH 7.5] (cat. no. HN77.1; Carl Roth) 12 mM MgCl2 (cat. no. HNO3.1; Carl Roth), 2 mM spermidine (cat. no. SO266-5G; Sigma-Aldrich), 40 mM dithiothreitol (DTT) (cat. no. 43915-5G; Sigma-Aldrich), a 3.125 mM concentration of each nucleoside triphosphate (cat. no. 11140957001, 11140922001, 11140965001, 11140949001; Roche, Basel, Switzerland), 1 U of RNasin/mL (cat. no. N2515; Promega, Mannheim, Germany), 0.1 μg of plasmid DNA/μL, and 0.6 U of T7 RNA polymerase/μL (cat. no. P2077; Promega).

    Produced:

    Article Title: Oxidized Low-Density Lipoprotein Is a Novel Predictor of Interferon Responsiveness in Chronic Hepatitis C Infection
    Article Snippet: Hepatitis C Virus RNA Transcription We produced HCV RNA by in vitro transcription, as described by Koutsoudakis et al. .. The in vitro transcription reaction mixture contained 80 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, pH 7.5] (cat. no. HN77.1; Carl Roth) 12 mM MgCl2 (cat. no. HNO3.1; Carl Roth), 2 mM spermidine (cat. no. SO266-5G; Sigma-Aldrich), 40 mM dithiothreitol (DTT) (cat. no. 43915-5G; Sigma-Aldrich), a 3.125 mM concentration of each nucleoside triphosphate (cat. no. 11140957001, 11140922001, 11140965001, 11140949001; Roche, Basel, Switzerland), 1 U of RNasin/mL (cat. no. N2515; Promega, Mannheim, Germany), 0.1 μg of plasmid DNA/μL, and 0.6 U of T7 RNA polymerase/μL (cat. no. P2077; Promega).

    Concentration Assay:

    Article Title: Oxidized Low-Density Lipoprotein Is a Novel Predictor of Interferon Responsiveness in Chronic Hepatitis C Infection
    Article Snippet: .. The in vitro transcription reaction mixture contained 80 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, pH 7.5] (cat. no. HN77.1; Carl Roth) 12 mM MgCl2 (cat. no. HNO3.1; Carl Roth), 2 mM spermidine (cat. no. SO266-5G; Sigma-Aldrich), 40 mM dithiothreitol (DTT) (cat. no. 43915-5G; Sigma-Aldrich), a 3.125 mM concentration of each nucleoside triphosphate (cat. no. 11140957001, 11140922001, 11140965001, 11140949001; Roche, Basel, Switzerland), 1 U of RNasin/mL (cat. no. N2515; Promega, Mannheim, Germany), 0.1 μg of plasmid DNA/μL, and 0.6 U of T7 RNA polymerase/μL (cat. no. P2077; Promega). .. After 2 hours of incubation at 37°C, 0.3 U T7 RNA polymerase was added to the mixture and incubated for another 2 hours at 37°C.

    Incubation:

    Article Title: Oxidized Low-Density Lipoprotein Is a Novel Predictor of Interferon Responsiveness in Chronic Hepatitis C Infection
    Article Snippet: The in vitro transcription reaction mixture contained 80 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, pH 7.5] (cat. no. HN77.1; Carl Roth) 12 mM MgCl2 (cat. no. HNO3.1; Carl Roth), 2 mM spermidine (cat. no. SO266-5G; Sigma-Aldrich), 40 mM dithiothreitol (DTT) (cat. no. 43915-5G; Sigma-Aldrich), a 3.125 mM concentration of each nucleoside triphosphate (cat. no. 11140957001, 11140922001, 11140965001, 11140949001; Roche, Basel, Switzerland), 1 U of RNasin/mL (cat. no. N2515; Promega, Mannheim, Germany), 0.1 μg of plasmid DNA/μL, and 0.6 U of T7 RNA polymerase/μL (cat. no. P2077; Promega). .. After 2 hours of incubation at 37°C, 0.3 U T7 RNA polymerase was added to the mixture and incubated for another 2 hours at 37°C.

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: Protein Identification Pipeline for the Homology Driven Proteomics
    Article Snippet: .. Cleland’s reagent (DTT) and iodoacetamide were of analytical grade and purchased from Sigma-Aldrich (Munich, Germany); water and acetonitrile were of LC-MS grade from Fisher Scientific (Schwerte, Germany). .. Formic acid and trifluoroacetic acid were HPLC grade obtained from Merck (Darmstadt, Germany).

    Sequencing:

    Article Title: Protein Identification Pipeline for the Homology Driven Proteomics
    Article Snippet: Cleland’s reagent (DTT) and iodoacetamide were of analytical grade and purchased from Sigma-Aldrich (Munich, Germany); water and acetonitrile were of LC-MS grade from Fisher Scientific (Schwerte, Germany). .. Modified porcine trypsin (sequencing grade) was purchased from Promega (Mannheim, Germany).

    Immunofluorescence:

    Article Title: ZAR1 and ZAR2 are required for oocyte meiotic maturation by regulating the maternal transcriptome and mRNA translational activation
    Article Snippet: .. Chromosome spreading and immunofluorescence Zona pellucida-free oocytes were fixed in a solution containing 1% paraformaldehyde, 0.15% Triton X-100 and 3 mmol/l dithiothreitol (DTT) (Sigma-Aldrich) on glass slides for 30 min and air dried. .. Cell culture, plasmid transfection and immunoprecipitation HeLa cells were obtained from American tissue culture collection and cultured in Dulbecco's-modified Eagle's medium (Invitrogen) added with 10% fetal bovine serum (FBS; Hyclone) and 1% penicillin-streptomycin solution (Gibco) at 37°C with 5% CO2 .

    Plasmid Preparation:

    Article Title: Oxidized Low-Density Lipoprotein Is a Novel Predictor of Interferon Responsiveness in Chronic Hepatitis C Infection
    Article Snippet: .. The in vitro transcription reaction mixture contained 80 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, pH 7.5] (cat. no. HN77.1; Carl Roth) 12 mM MgCl2 (cat. no. HNO3.1; Carl Roth), 2 mM spermidine (cat. no. SO266-5G; Sigma-Aldrich), 40 mM dithiothreitol (DTT) (cat. no. 43915-5G; Sigma-Aldrich), a 3.125 mM concentration of each nucleoside triphosphate (cat. no. 11140957001, 11140922001, 11140965001, 11140949001; Roche, Basel, Switzerland), 1 U of RNasin/mL (cat. no. N2515; Promega, Mannheim, Germany), 0.1 μg of plasmid DNA/μL, and 0.6 U of T7 RNA polymerase/μL (cat. no. P2077; Promega). .. After 2 hours of incubation at 37°C, 0.3 U T7 RNA polymerase was added to the mixture and incubated for another 2 hours at 37°C.

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    Millipore dtt
    Shown are 600 MHz 1 H- 15 N HSQC spectra of Parkin IBR 307–384 . ( a ) Folded 0.2 mM IBR 307–384 in 25 mM phosphate buffer (pH 6.95), 100 mM <t>NaCl,</t> 1 mM <t>DTT</t> at 25°C. ( b ) Spectrum of IBR 307–384 from a upon EDTA addition showing the
    Dtt, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1308 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dtt/product/Millipore
    Average 95 stars, based on 1308 article reviews
    Price from $9.99 to $1999.99
    dtt - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    78
    Millipore uch buffer
    Insoluble α-syn in diseased brain is ubiquitinated. A: Western blot analysis of biochemically fractionated cingulate cortex from a patient with <t>DLB</t> (DLB-3). Immunoblots were developed with anti-α-syn antibodies LB509 and Syn208 as well as anti-ubiquitin antibody mAb 1510. Twenty μl of LS fraction ( lane 1 ), TX fraction ( lane 2 ), sarkosyl-soluble fraction ( lane 3 ), and SDS-soluble fraction ( lane 4 ) were loaded in separate lanes of 15% SDS-polyacrylamide gels. Note that the SDS-soluble fraction is four times as concentrated as each of the other fractions in that 2.5 ml/g of SDS buffer was used for tissue extraction versus 10 ml/g for each of the other fractions. Arrowhead indicates α-syn monomer (αS) and bracket indicates ubiquitin monomer (Ub). Arrows depict mono-, di-, and tri-ubiquitinated forms of α-syn. B: Western blot analysis (with antibodies Syn208 and mAb 1510) of the SDS-soluble fraction from the cingulate cortex of normal brains (NL-1, NL-2) and DLB brains (DLB-1, DLB-2, and DLB-3). Twenty μl of SDS-soluble fraction was loaded in each lane of a 15% gel. One hundred ng of recombinant human α-syn was loaded in the indicated lanes. C: Western blot analysis of the SDS-soluble fraction from the cingulate cortex of case DLB-1 using various anti-α-syn (LB509, Syn102, Syn211) and anti-ubiquitin (mAb 1510, Conj8) antibodies. Arrows depict mono-, di-, and tri-ubiquitinated forms of α-syn. D: Immunoprecipitation followed by Western blot analysis. α-Syn in the SDS-soluble fraction of the cingulate cortex of DLB-1 was isolated by immunoprecipitation with anti-α-syn antibodies. The sample was analyzed by Western blot analysis using anti-α-syn antibody LB509 and anti-ubiquitin antibody mAb 1510 ( arrows , mono- and di-ubiquitinated forms of α-syn; * and **, possible ubiquitinated forms of α-syn in which ubiquitin moieties may be masking the LB509 epitope; ***, modified form of α-syn, possibly dimerized α-syn). E: Ubiquitinated α-syn from DLB brain can be deubiquitinated by <t>UCH-L1</t> in vitro . SDS-soluble fraction from the cingulate cortex of NL-1 or DLB-1 was untreated or reacted with 50 nmol/L of UCH-L1. The samples were analyzed by Western blot analysis using LB509. R represents a lane loaded with 100 ng of recombinant human α-syn. The mobility of molecular mass markers (kd) is depicted on the left of each panel.
    Uch Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/uch buffer/product/Millipore
    Average 78 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    uch buffer - by Bioz Stars, 2020-02
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    95
    Millipore ubiquitination buffer
    Function of PDLIM2 phosphorylation mutants, S137A, T138A, and S137D, in in vivo and in vitro <t>ubiquitination</t> assays. a, in vitro ubiquitination assay. Recombinant His-tagged PDLIM2, His-tagged PDLIM2 mutants, or empty His vector were transfected and expressed
    Ubiquitination Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ubiquitination buffer/product/Millipore
    Average 95 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    ubiquitination buffer - by Bioz Stars, 2020-02
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    Image Search Results


    Shown are 600 MHz 1 H- 15 N HSQC spectra of Parkin IBR 307–384 . ( a ) Folded 0.2 mM IBR 307–384 in 25 mM phosphate buffer (pH 6.95), 100 mM NaCl, 1 mM DTT at 25°C. ( b ) Spectrum of IBR 307–384 from a upon EDTA addition showing the

    Journal:

    Article Title: Structure of the Parkin in-between-ring domain provides insights for E3-ligase dysfunction in autosomal recessive Parkinson's disease

    doi: 10.1073/pnas.0610548104

    Figure Lengend Snippet: Shown are 600 MHz 1 H- 15 N HSQC spectra of Parkin IBR 307–384 . ( a ) Folded 0.2 mM IBR 307–384 in 25 mM phosphate buffer (pH 6.95), 100 mM NaCl, 1 mM DTT at 25°C. ( b ) Spectrum of IBR 307–384 from a upon EDTA addition showing the

    Article Snippet: Proteins were dialyzed against 25 mM Na2 HPO4 pH 6.95, 100 mM NaCl, and 1 mM DTT and concentrated to 0.2–0.9 mM by ultrafiltration (Millipore, Mississauga, ON, Canada) for NMR experiments.

    Techniques:

    Insoluble α-syn in diseased brain is ubiquitinated. A: Western blot analysis of biochemically fractionated cingulate cortex from a patient with DLB (DLB-3). Immunoblots were developed with anti-α-syn antibodies LB509 and Syn208 as well as anti-ubiquitin antibody mAb 1510. Twenty μl of LS fraction ( lane 1 ), TX fraction ( lane 2 ), sarkosyl-soluble fraction ( lane 3 ), and SDS-soluble fraction ( lane 4 ) were loaded in separate lanes of 15% SDS-polyacrylamide gels. Note that the SDS-soluble fraction is four times as concentrated as each of the other fractions in that 2.5 ml/g of SDS buffer was used for tissue extraction versus 10 ml/g for each of the other fractions. Arrowhead indicates α-syn monomer (αS) and bracket indicates ubiquitin monomer (Ub). Arrows depict mono-, di-, and tri-ubiquitinated forms of α-syn. B: Western blot analysis (with antibodies Syn208 and mAb 1510) of the SDS-soluble fraction from the cingulate cortex of normal brains (NL-1, NL-2) and DLB brains (DLB-1, DLB-2, and DLB-3). Twenty μl of SDS-soluble fraction was loaded in each lane of a 15% gel. One hundred ng of recombinant human α-syn was loaded in the indicated lanes. C: Western blot analysis of the SDS-soluble fraction from the cingulate cortex of case DLB-1 using various anti-α-syn (LB509, Syn102, Syn211) and anti-ubiquitin (mAb 1510, Conj8) antibodies. Arrows depict mono-, di-, and tri-ubiquitinated forms of α-syn. D: Immunoprecipitation followed by Western blot analysis. α-Syn in the SDS-soluble fraction of the cingulate cortex of DLB-1 was isolated by immunoprecipitation with anti-α-syn antibodies. The sample was analyzed by Western blot analysis using anti-α-syn antibody LB509 and anti-ubiquitin antibody mAb 1510 ( arrows , mono- and di-ubiquitinated forms of α-syn; * and **, possible ubiquitinated forms of α-syn in which ubiquitin moieties may be masking the LB509 epitope; ***, modified form of α-syn, possibly dimerized α-syn). E: Ubiquitinated α-syn from DLB brain can be deubiquitinated by UCH-L1 in vitro . SDS-soluble fraction from the cingulate cortex of NL-1 or DLB-1 was untreated or reacted with 50 nmol/L of UCH-L1. The samples were analyzed by Western blot analysis using LB509. R represents a lane loaded with 100 ng of recombinant human α-syn. The mobility of molecular mass markers (kd) is depicted on the left of each panel.

    Journal: The American Journal of Pathology

    Article Title: Ubiquitination of ?-Synuclein Is Not Required for Formation of Pathological Inclusions in ?-Synucleinopathies

    doi:

    Figure Lengend Snippet: Insoluble α-syn in diseased brain is ubiquitinated. A: Western blot analysis of biochemically fractionated cingulate cortex from a patient with DLB (DLB-3). Immunoblots were developed with anti-α-syn antibodies LB509 and Syn208 as well as anti-ubiquitin antibody mAb 1510. Twenty μl of LS fraction ( lane 1 ), TX fraction ( lane 2 ), sarkosyl-soluble fraction ( lane 3 ), and SDS-soluble fraction ( lane 4 ) were loaded in separate lanes of 15% SDS-polyacrylamide gels. Note that the SDS-soluble fraction is four times as concentrated as each of the other fractions in that 2.5 ml/g of SDS buffer was used for tissue extraction versus 10 ml/g for each of the other fractions. Arrowhead indicates α-syn monomer (αS) and bracket indicates ubiquitin monomer (Ub). Arrows depict mono-, di-, and tri-ubiquitinated forms of α-syn. B: Western blot analysis (with antibodies Syn208 and mAb 1510) of the SDS-soluble fraction from the cingulate cortex of normal brains (NL-1, NL-2) and DLB brains (DLB-1, DLB-2, and DLB-3). Twenty μl of SDS-soluble fraction was loaded in each lane of a 15% gel. One hundred ng of recombinant human α-syn was loaded in the indicated lanes. C: Western blot analysis of the SDS-soluble fraction from the cingulate cortex of case DLB-1 using various anti-α-syn (LB509, Syn102, Syn211) and anti-ubiquitin (mAb 1510, Conj8) antibodies. Arrows depict mono-, di-, and tri-ubiquitinated forms of α-syn. D: Immunoprecipitation followed by Western blot analysis. α-Syn in the SDS-soluble fraction of the cingulate cortex of DLB-1 was isolated by immunoprecipitation with anti-α-syn antibodies. The sample was analyzed by Western blot analysis using anti-α-syn antibody LB509 and anti-ubiquitin antibody mAb 1510 ( arrows , mono- and di-ubiquitinated forms of α-syn; * and **, possible ubiquitinated forms of α-syn in which ubiquitin moieties may be masking the LB509 epitope; ***, modified form of α-syn, possibly dimerized α-syn). E: Ubiquitinated α-syn from DLB brain can be deubiquitinated by UCH-L1 in vitro . SDS-soluble fraction from the cingulate cortex of NL-1 or DLB-1 was untreated or reacted with 50 nmol/L of UCH-L1. The samples were analyzed by Western blot analysis using LB509. R represents a lane loaded with 100 ng of recombinant human α-syn. The mobility of molecular mass markers (kd) is depicted on the left of each panel.

    Article Snippet: SDS-soluble fractions from the cingulate cortex of neuropathologically normal (NL) or DLB brains were diluted 40-fold in UCH buffer (50 mmol/L HEPES, pH 7.8, 0.5 mmol/L EDTA, 1 mmol/L dithiothreitol) and then concentrated 40-fold using a MicroconYM-10 (Millipore Corp., Bedford, MA) to remove SDS.

    Techniques: Western Blot, Recombinant, Immunoprecipitation, Isolation, Modification, In Vitro

    Function of PDLIM2 phosphorylation mutants, S137A, T138A, and S137D, in in vivo and in vitro ubiquitination assays. a, in vitro ubiquitination assay. Recombinant His-tagged PDLIM2, His-tagged PDLIM2 mutants, or empty His vector were transfected and expressed

    Journal: The Journal of Biological Chemistry

    Article Title: Osteopontin and Protein Kinase C Regulate PDLIM2 Activation and STAT1 Ubiquitination in LPS-treated Murine Macrophages *

    doi: 10.1074/jbc.M110.161869

    Figure Lengend Snippet: Function of PDLIM2 phosphorylation mutants, S137A, T138A, and S137D, in in vivo and in vitro ubiquitination assays. a, in vitro ubiquitination assay. Recombinant His-tagged PDLIM2, His-tagged PDLIM2 mutants, or empty His vector were transfected and expressed

    Article Snippet: Anti-FLAG STAT1 immunoprecipitates were washed five times with lysis buffer and once with ubiquitination buffer (50 m m Tris-Cl, pH 7.5, 2 m m MgCl2 , 2 m m ATP, and 1 m m DTT) and incubated in 40 μl of ubiquitination buffer with 5 μg of Ub, 100 ng of Ub-activating enzyme (E1), and 100 ng of GST-UbcH5a (Calbiochem) in the absence or presence of recombinant PDLIM2 for 3 h at 30 °C.

    Techniques: In Vivo, In Vitro, Ubiquitin Assay, Recombinant, Plasmid Preparation, Transfection