1 4 dithiothreitol  (Thermo Fisher)


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    Structured Review

    Thermo Fisher 1 4 dithiothreitol
    1 4 Dithiothreitol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1 4 dithiothreitol/product/Thermo Fisher
    Average 99 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    1 4 dithiothreitol - by Bioz Stars, 2020-03
    99/100 stars

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    Centrifugation:

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: Western blotting analysis Cells were lysed in lysis buffer [0.5% Nonidet P-40 (IGEPAL® CA-630 Sigma-Aldrich I3021), 137 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 4 mM sodium orthovanadate, 20 mM glycerol-2-phosphate, 10 mM sodium fluoride, 1 mM sodium pyrophosphate], together with complete protease and Phospho-STOP inhibitor mixtures (Roche Applied Science, 05 892 970 001) for 30 min on ice and cleared by centrifugation for 10 min at 20,817 g force. .. After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290).

    Amplification:

    Article Title: Inheritance of susceptibility to induction of nephroblastomas in the Noble rat
    Article Snippet: Briefly, RNA samples were reverse transcribed at 42° C for 1 hr by incubation with 20 μl of a reverse transcription mixture containing the following components: 5 μM of hexamer primer; 1X reverse transcriptase buffer; 10 mM 1,4-dithiothreitol; 5 mM MgCl2 ; 20 units of RNasin; 0.5 mM deoxynucleotide triphosphates; 200 units of SuperScript II (Invitrogen, CA). .. Repeated PCR reactions for each primer pair were performed to optimize the number of cycles required to remain within the exponential phase of the amplification curve (data not shown).

    Article Title: A Subset of Odorant Receptors from the Desert Locust Schistocerca gregaria Is Co-Expressed with the Sensory Neuron Membrane Protein 1
    Article Snippet: Following a 5-min incubation at 65 °C, 8 µL 5× SSIV Buffer (Thermo Fisher Scientific), 2 μL 100 mM 1,4-dithiothreitol (DTT; Thermo Fisher Scientific), 2 µL RNaseOut (Thermo Fisher Scientific) and 1 µL Superscript IV Reverse Transcriptase (Thermo Fisher Scientific) were added. .. For the subsequent amplification of sequences encoding actin (GenBank accession number: AEV89776) or the OR types SgreOR84-86, SgreOR88, SgreOR90-97, SgreOR101-103, SgreOR109-114, SgreOR116 and SgreOR118 in PCR experiments, the oligonucleotide primers given in were utilized.

    Synthesized:

    Article Title: Structure-Guided Immobilization of an Evolved Unspecific Peroxygenase
    Article Snippet: .. The oligonucleotides were synthesized by Isogen Life Science (De Meern, The Netherlands), and NBD, naphthalene, styrene, 2,2’-dithiodipyridine and 1,4-dithiothreitol were purchased from Acros Organics (Waltham, MA, USA). ..

    Blocking Assay:

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290). .. Blocking of the membrane was performed using 5% milk powder diluted in PBS (ThermoFisher Scientific, 14190250), supplemented with 0.1% Tween 20 (ThermoFisher Scientific, 28352).

    Real-time Polymerase Chain Reaction:

    Article Title: Impact of LytR-CpsA-Psr Proteins on Cell Wall Biosynthesis in Corynebacterium glutamicum
    Article Snippet: The samples were immediately cooled on ice and then mixed with the following solutions: 4 μl first-strand buffer (5×), 2 μl 1,4-dithiothreitol (DTT; 0.1 M), 1 μl deoxynucleoside triphosphates (10 mM each), 2 μl SuperScript III reverse transcriptase (200 U/μl; Life Technologies, Darmstadt, Germany). .. The samples were mixed with 179 μl H2 O to reach a concentration of about 5 ng/μl cDNA for the qPCR. qPCR was performed in a qTOWER real-time PCR cycler (Analytic Jena, Jena, Germany) using innu Mix qPCR master mix SyGreen (Analytic Jena, Jena, Germany).

    Microarray:

    Article Title: Inheritance of susceptibility to induction of nephroblastomas in the Noble rat
    Article Snippet: For semi-quantitative RT-PCR confirmation of microarray results, total RNA (1 μg) was used for the synthesis of single-stranded cDNA. .. Briefly, RNA samples were reverse transcribed at 42° C for 1 hr by incubation with 20 μl of a reverse transcription mixture containing the following components: 5 μM of hexamer primer; 1X reverse transcriptase buffer; 10 mM 1,4-dithiothreitol; 5 mM MgCl2 ; 20 units of RNasin; 0.5 mM deoxynucleotide triphosphates; 200 units of SuperScript II (Invitrogen, CA).

    Incubation:

    Article Title: Inheritance of susceptibility to induction of nephroblastomas in the Noble rat
    Article Snippet: .. Briefly, RNA samples were reverse transcribed at 42° C for 1 hr by incubation with 20 μl of a reverse transcription mixture containing the following components: 5 μM of hexamer primer; 1X reverse transcriptase buffer; 10 mM 1,4-dithiothreitol; 5 mM MgCl2 ; 20 units of RNasin; 0.5 mM deoxynucleotide triphosphates; 200 units of SuperScript II (Invitrogen, CA). .. The resulting single-stranded cDNA was used for PCR.

    Article Title: ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function
    Article Snippet: Lysates were partly harvested and diluted in loading buffer complemented with 250 mM 1,4-dithiothreitol (DTT; Thermo Scientific). .. The lysates were pre-cleaned through incubation with Dynabeads Protein G for IP (Life Technologies) at 4 °C for 2 hours.

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290). .. Afterwards, membranes were incubated with a primary antibody overnight at 4°C and with a secondary horseradish peroxidase conjugated antibody (Jackson ImmunoResearch Europe Ltd, goat anti-mouse: 115-035-003, goat anti-rabbit: 111-035-144) for 1 h. Blots were developed by using the enhanced chemiluminescence ECL PlusTM detection system (ThermoFisher Scientific, 32134) and imaged with an Amersham 600 Imager operated with ImageQuant software (GE Healthcare Life Sciences) and using the automated exposure program.

    Article Title: Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B12 Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis ▿ ▿ †
    Article Snippet: .. In brief, two 50-μl reaction mixtures containing 1× transcription buffer (40 mM Tris-HCl, pH 8, 10 mM MgCl2 , 0.1 mM EDTA, pH 8, 50 mM KCl), 70 nM PCR template, 1× initiation nucleoside triphosphates (NTPs) (1 μM ATP, 1 μM GTP, 0.4 μM UTP [lithium salt; Roche Applied Science]), 10 μg/ml bovine serum albumin, 1 mM 1-4-dithiothreitol, 30 U RNaseOut (Invitrogen), 66 nM [α-32 P]UTP (3,000 Ci/mmol, 10 mCi/ml) (Perkin Elmer), and 68 nM E. coli RNA polymerase (Epicentre) were set up for each template and incubated for 10 min at 37°C. .. Elongation was initiated by immediately adding 6.25 μl of 50 mM CoB12 or diethylpyrocarbonate (DEPC)-treated water and 6.25 μl of elongation mix (60 μl of elongation mix contained 6 μl of 10× transcription buffer, 12 μl of 20 mg/ml heparin [grade 1-A; Sigma], 6 μl each of 1 mM ATP, CTP, GTP, and UTP, and 18 μl of DEPC-treated water).

    Article Title: A Subset of Odorant Receptors from the Desert Locust Schistocerca gregaria Is Co-Expressed with the Sensory Neuron Membrane Protein 1
    Article Snippet: .. Following a 5-min incubation at 65 °C, 8 µL 5× SSIV Buffer (Thermo Fisher Scientific), 2 μL 100 mM 1,4-dithiothreitol (DTT; Thermo Fisher Scientific), 2 µL RNaseOut (Thermo Fisher Scientific) and 1 µL Superscript IV Reverse Transcriptase (Thermo Fisher Scientific) were added. ..

    Article Title: Impact of LytR-CpsA-Psr Proteins on Cell Wall Biosynthesis in Corynebacterium glutamicum
    Article Snippet: For reverse transcription, 1 μg RNA in a total volume of 10 μl was mixed with 1 μl random primers (0.5 μg/μl; Life Technologies, Darmstadt, Germany) and incubated at 70°C for 10 min. .. The samples were immediately cooled on ice and then mixed with the following solutions: 4 μl first-strand buffer (5×), 2 μl 1,4-dithiothreitol (DTT; 0.1 M), 1 μl deoxynucleoside triphosphates (10 mM each), 2 μl SuperScript III reverse transcriptase (200 U/μl; Life Technologies, Darmstadt, Germany).

    Expressing:

    Article Title: A Subset of Odorant Receptors from the Desert Locust Schistocerca gregaria Is Co-Expressed with the Sensory Neuron Membrane Protein 1
    Article Snippet: Paragraph title: 2.6. RNA Extraction and Reverse Transcription PCR for Comparing SgreOR Expression in Males Versus Females ... Following a 5-min incubation at 65 °C, 8 µL 5× SSIV Buffer (Thermo Fisher Scientific), 2 μL 100 mM 1,4-dithiothreitol (DTT; Thermo Fisher Scientific), 2 µL RNaseOut (Thermo Fisher Scientific) and 1 µL Superscript IV Reverse Transcriptase (Thermo Fisher Scientific) were added.

    BIA-KA:

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: .. After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290). .. Proteins were separated on NuPAGE gels (Life Technologies, NP0322BOX) or on 18% gels made in-house (for detection of LC3 isoforms) and transferred to a nitrocellulose membrane HybondTM -P (GE Healthcare Life Sciences, RPN132D).

    Western Blot:

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: Paragraph title: Western blotting analysis ... After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290).

    Transformation Assay:

    Article Title: Structure-Guided Immobilization of an Evolved Unspecific Peroxygenase
    Article Snippet: Reagents and Enzymes ABTS, hemoglobin from bovine blood, the S. cerevisiae transformation kit, the activated Thiol–Sepharose® and fluoresecein isothiocyanate were all purchased from Sigma-Aldrich/Merck (Saint Louis, MO, USA). .. The oligonucleotides were synthesized by Isogen Life Science (De Meern, The Netherlands), and NBD, naphthalene, styrene, 2,2’-dithiodipyridine and 1,4-dithiothreitol were purchased from Acros Organics (Waltham, MA, USA).

    Transfection:

    Article Title: ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function
    Article Snippet: Additional controls untransfected and transfected with either TBK1 WT or OPTN WT only were used. .. Lysates were partly harvested and diluted in loading buffer complemented with 250 mM 1,4-dithiothreitol (DTT; Thermo Scientific).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Inheritance of susceptibility to induction of nephroblastomas in the Noble rat
    Article Snippet: Paragraph title: RT-PCR and Sequencing ... Briefly, RNA samples were reverse transcribed at 42° C for 1 hr by incubation with 20 μl of a reverse transcription mixture containing the following components: 5 μM of hexamer primer; 1X reverse transcriptase buffer; 10 mM 1,4-dithiothreitol; 5 mM MgCl2 ; 20 units of RNasin; 0.5 mM deoxynucleotide triphosphates; 200 units of SuperScript II (Invitrogen, CA).

    Generated:

    Article Title: Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B12 Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis ▿ ▿ †
    Article Snippet: Templates (P2, SP2, SP3, and SP4) ( ) for the in vitro termination assays were generated by PCR with primers listed in Tables and . .. In brief, two 50-μl reaction mixtures containing 1× transcription buffer (40 mM Tris-HCl, pH 8, 10 mM MgCl2 , 0.1 mM EDTA, pH 8, 50 mM KCl), 70 nM PCR template, 1× initiation nucleoside triphosphates (NTPs) (1 μM ATP, 1 μM GTP, 0.4 μM UTP [lithium salt; Roche Applied Science]), 10 μg/ml bovine serum albumin, 1 mM 1-4-dithiothreitol, 30 U RNaseOut (Invitrogen), 66 nM [α-32 P]UTP (3,000 Ci/mmol, 10 mCi/ml) (Perkin Elmer), and 68 nM E. coli RNA polymerase (Epicentre) were set up for each template and incubated for 10 min at 37°C.

    Protein Concentration:

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: .. After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290). .. Proteins were separated on NuPAGE gels (Life Technologies, NP0322BOX) or on 18% gels made in-house (for detection of LC3 isoforms) and transferred to a nitrocellulose membrane HybondTM -P (GE Healthcare Life Sciences, RPN132D).

    Sequencing:

    Article Title: Inheritance of susceptibility to induction of nephroblastomas in the Noble rat
    Article Snippet: Paragraph title: RT-PCR and Sequencing ... Briefly, RNA samples were reverse transcribed at 42° C for 1 hr by incubation with 20 μl of a reverse transcription mixture containing the following components: 5 μM of hexamer primer; 1X reverse transcriptase buffer; 10 mM 1,4-dithiothreitol; 5 mM MgCl2 ; 20 units of RNasin; 0.5 mM deoxynucleotide triphosphates; 200 units of SuperScript II (Invitrogen, CA).

    Binding Assay:

    Article Title: ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function
    Article Snippet: Lysates were partly harvested and diluted in loading buffer complemented with 250 mM 1,4-dithiothreitol (DTT; Thermo Scientific). .. As an additional negative control, 2 samples transfected with OPTN WT only or TBK1 WT only were incubated with beads and no antibody, to reveal any unspecific binding.

    Nucleic Acid Electrophoresis:

    Article Title: Impact of LytR-CpsA-Psr Proteins on Cell Wall Biosynthesis in Corynebacterium glutamicum
    Article Snippet: Afterwards, the RNA was purified by phenol-chloroform extraction and resuspended in 15 μl RNase-free H2 O. RNA integrity was assessed using gel electrophoresis. .. The samples were immediately cooled on ice and then mixed with the following solutions: 4 μl first-strand buffer (5×), 2 μl 1,4-dithiothreitol (DTT; 0.1 M), 1 μl deoxynucleoside triphosphates (10 mM each), 2 μl SuperScript III reverse transcriptase (200 U/μl; Life Technologies, Darmstadt, Germany).

    Mutagenesis:

    Article Title: ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function
    Article Snippet: 2.11 Cotransfection IP assay Cells were transfected with TBK1 WT and mutant plasmids together with OPTN WT plasmids. .. Lysates were partly harvested and diluted in loading buffer complemented with 250 mM 1,4-dithiothreitol (DTT; Thermo Scientific).

    Negative Control:

    Article Title: ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function
    Article Snippet: Lysates were partly harvested and diluted in loading buffer complemented with 250 mM 1,4-dithiothreitol (DTT; Thermo Scientific). .. As an additional negative control, 2 samples transfected with OPTN WT only or TBK1 WT only were incubated with beads and no antibody, to reveal any unspecific binding.

    Purification:

    Article Title: Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B12 Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis ▿ ▿ †
    Article Snippet: The PCR products were purified using a QIAquick PCR purification kit (Qiagen). .. In brief, two 50-μl reaction mixtures containing 1× transcription buffer (40 mM Tris-HCl, pH 8, 10 mM MgCl2 , 0.1 mM EDTA, pH 8, 50 mM KCl), 70 nM PCR template, 1× initiation nucleoside triphosphates (NTPs) (1 μM ATP, 1 μM GTP, 0.4 μM UTP [lithium salt; Roche Applied Science]), 10 μg/ml bovine serum albumin, 1 mM 1-4-dithiothreitol, 30 U RNaseOut (Invitrogen), 66 nM [α-32 P]UTP (3,000 Ci/mmol, 10 mCi/ml) (Perkin Elmer), and 68 nM E. coli RNA polymerase (Epicentre) were set up for each template and incubated for 10 min at 37°C.

    Article Title: A Subset of Odorant Receptors from the Desert Locust Schistocerca gregaria Is Co-Expressed with the Sensory Neuron Membrane Protein 1
    Article Snippet: Next, poly(A)+ RNA was extracted with the Dynabeads mRNA purification kit following the manufacturer’s protocol. .. Following a 5-min incubation at 65 °C, 8 µL 5× SSIV Buffer (Thermo Fisher Scientific), 2 μL 100 mM 1,4-dithiothreitol (DTT; Thermo Fisher Scientific), 2 µL RNaseOut (Thermo Fisher Scientific) and 1 µL Superscript IV Reverse Transcriptase (Thermo Fisher Scientific) were added.

    Article Title: Impact of LytR-CpsA-Psr Proteins on Cell Wall Biosynthesis in Corynebacterium glutamicum
    Article Snippet: Afterwards, the RNA was purified by phenol-chloroform extraction and resuspended in 15 μl RNase-free H2 O. RNA integrity was assessed using gel electrophoresis. .. The samples were immediately cooled on ice and then mixed with the following solutions: 4 μl first-strand buffer (5×), 2 μl 1,4-dithiothreitol (DTT; 0.1 M), 1 μl deoxynucleoside triphosphates (10 mM each), 2 μl SuperScript III reverse transcriptase (200 U/μl; Life Technologies, Darmstadt, Germany).

    Polymerase Chain Reaction:

    Article Title: Inheritance of susceptibility to induction of nephroblastomas in the Noble rat
    Article Snippet: Briefly, RNA samples were reverse transcribed at 42° C for 1 hr by incubation with 20 μl of a reverse transcription mixture containing the following components: 5 μM of hexamer primer; 1X reverse transcriptase buffer; 10 mM 1,4-dithiothreitol; 5 mM MgCl2 ; 20 units of RNasin; 0.5 mM deoxynucleotide triphosphates; 200 units of SuperScript II (Invitrogen, CA). .. The resulting single-stranded cDNA was used for PCR.

    Article Title: Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B12 Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis ▿ ▿ †
    Article Snippet: .. In brief, two 50-μl reaction mixtures containing 1× transcription buffer (40 mM Tris-HCl, pH 8, 10 mM MgCl2 , 0.1 mM EDTA, pH 8, 50 mM KCl), 70 nM PCR template, 1× initiation nucleoside triphosphates (NTPs) (1 μM ATP, 1 μM GTP, 0.4 μM UTP [lithium salt; Roche Applied Science]), 10 μg/ml bovine serum albumin, 1 mM 1-4-dithiothreitol, 30 U RNaseOut (Invitrogen), 66 nM [α-32 P]UTP (3,000 Ci/mmol, 10 mCi/ml) (Perkin Elmer), and 68 nM E. coli RNA polymerase (Epicentre) were set up for each template and incubated for 10 min at 37°C. .. Elongation was initiated by immediately adding 6.25 μl of 50 mM CoB12 or diethylpyrocarbonate (DEPC)-treated water and 6.25 μl of elongation mix (60 μl of elongation mix contained 6 μl of 10× transcription buffer, 12 μl of 20 mg/ml heparin [grade 1-A; Sigma], 6 μl each of 1 mM ATP, CTP, GTP, and UTP, and 18 μl of DEPC-treated water).

    Article Title: A Subset of Odorant Receptors from the Desert Locust Schistocerca gregaria Is Co-Expressed with the Sensory Neuron Membrane Protein 1
    Article Snippet: Paragraph title: 2.6. RNA Extraction and Reverse Transcription PCR for Comparing SgreOR Expression in Males Versus Females ... Following a 5-min incubation at 65 °C, 8 µL 5× SSIV Buffer (Thermo Fisher Scientific), 2 μL 100 mM 1,4-dithiothreitol (DTT; Thermo Fisher Scientific), 2 µL RNaseOut (Thermo Fisher Scientific) and 1 µL Superscript IV Reverse Transcriptase (Thermo Fisher Scientific) were added.

    Quantitative RT-PCR:

    Article Title: Impact of LytR-CpsA-Psr Proteins on Cell Wall Biosynthesis in Corynebacterium glutamicum
    Article Snippet: Paragraph title: RT-qPCR. ... The samples were immediately cooled on ice and then mixed with the following solutions: 4 μl first-strand buffer (5×), 2 μl 1,4-dithiothreitol (DTT; 0.1 M), 1 μl deoxynucleoside triphosphates (10 mM each), 2 μl SuperScript III reverse transcriptase (200 U/μl; Life Technologies, Darmstadt, Germany).

    Cotransfection:

    Article Title: ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function
    Article Snippet: Paragraph title: Cotransfection IP assay ... Lysates were partly harvested and diluted in loading buffer complemented with 250 mM 1,4-dithiothreitol (DTT; Thermo Scientific).

    Plasmid Preparation:

    Article Title: Structure-Guided Immobilization of an Evolved Unspecific Peroxygenase
    Article Snippet: The uracil independent and ampicillin resistance shuttle vector pJRoC30 was obtained from the California Institute of Technology (CALTECH, Pasadena, CA, USA). .. The oligonucleotides were synthesized by Isogen Life Science (De Meern, The Netherlands), and NBD, naphthalene, styrene, 2,2’-dithiodipyridine and 1,4-dithiothreitol were purchased from Acros Organics (Waltham, MA, USA).

    Software:

    Article Title: Inheritance of susceptibility to induction of nephroblastomas in the Noble rat
    Article Snippet: Briefly, RNA samples were reverse transcribed at 42° C for 1 hr by incubation with 20 μl of a reverse transcription mixture containing the following components: 5 μM of hexamer primer; 1X reverse transcriptase buffer; 10 mM 1,4-dithiothreitol; 5 mM MgCl2 ; 20 units of RNasin; 0.5 mM deoxynucleotide triphosphates; 200 units of SuperScript II (Invitrogen, CA). .. The oligonucleotide primers for selected genes were designed using GeneFisher software (sequences are available upon request).

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290). .. Afterwards, membranes were incubated with a primary antibody overnight at 4°C and with a secondary horseradish peroxidase conjugated antibody (Jackson ImmunoResearch Europe Ltd, goat anti-mouse: 115-035-003, goat anti-rabbit: 111-035-144) for 1 h. Blots were developed by using the enhanced chemiluminescence ECL PlusTM detection system (ThermoFisher Scientific, 32134) and imaged with an Amersham 600 Imager operated with ImageQuant software (GE Healthcare Life Sciences) and using the automated exposure program.

    RNA Extraction:

    Article Title: A Subset of Odorant Receptors from the Desert Locust Schistocerca gregaria Is Co-Expressed with the Sensory Neuron Membrane Protein 1
    Article Snippet: Paragraph title: 2.6. RNA Extraction and Reverse Transcription PCR for Comparing SgreOR Expression in Males Versus Females ... Following a 5-min incubation at 65 °C, 8 µL 5× SSIV Buffer (Thermo Fisher Scientific), 2 μL 100 mM 1,4-dithiothreitol (DTT; Thermo Fisher Scientific), 2 µL RNaseOut (Thermo Fisher Scientific) and 1 µL Superscript IV Reverse Transcriptase (Thermo Fisher Scientific) were added.

    In Vitro:

    Article Title: Transcription Antitermination by a Phosphorylated Response Regulator and Cobalamin-Dependent Termination at a B12 Riboswitch Contribute to Ethanolamine Utilization in Enterococcus faecalis ▿ ▿ †
    Article Snippet: Paragraph title: In vitro termination assays with CoB12. ... In brief, two 50-μl reaction mixtures containing 1× transcription buffer (40 mM Tris-HCl, pH 8, 10 mM MgCl2 , 0.1 mM EDTA, pH 8, 50 mM KCl), 70 nM PCR template, 1× initiation nucleoside triphosphates (NTPs) (1 μM ATP, 1 μM GTP, 0.4 μM UTP [lithium salt; Roche Applied Science]), 10 μg/ml bovine serum albumin, 1 mM 1-4-dithiothreitol, 30 U RNaseOut (Invitrogen), 66 nM [α-32 P]UTP (3,000 Ci/mmol, 10 mCi/ml) (Perkin Elmer), and 68 nM E. coli RNA polymerase (Epicentre) were set up for each template and incubated for 10 min at 37°C.

    Spectrophotometry:

    Article Title: A Subset of Odorant Receptors from the Desert Locust Schistocerca gregaria Is Co-Expressed with the Sensory Neuron Membrane Protein 1
    Article Snippet: Finally, poly(A)+ RNA samples were eluted with 22 µl H2 O and RNA concentrations were measured by means of an Epoch microplate spectrophotometer (BioTek, Winooski, VT, USA). .. Following a 5-min incubation at 65 °C, 8 µL 5× SSIV Buffer (Thermo Fisher Scientific), 2 μL 100 mM 1,4-dithiothreitol (DTT; Thermo Fisher Scientific), 2 µL RNaseOut (Thermo Fisher Scientific) and 1 µL Superscript IV Reverse Transcriptase (Thermo Fisher Scientific) were added.

    Concentration Assay:

    Article Title: Impact of LytR-CpsA-Psr Proteins on Cell Wall Biosynthesis in Corynebacterium glutamicum
    Article Snippet: The samples were immediately cooled on ice and then mixed with the following solutions: 4 μl first-strand buffer (5×), 2 μl 1,4-dithiothreitol (DTT; 0.1 M), 1 μl deoxynucleoside triphosphates (10 mM each), 2 μl SuperScript III reverse transcriptase (200 U/μl; Life Technologies, Darmstadt, Germany). .. The samples were mixed with 179 μl H2 O to reach a concentration of about 5 ng/μl cDNA for the qPCR. qPCR was performed in a qTOWER real-time PCR cycler (Analytic Jena, Jena, Germany) using innu Mix qPCR master mix SyGreen (Analytic Jena, Jena, Germany).

    Lysis:

    Article Title: Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1/p62 bodies
    Article Snippet: Western blotting analysis Cells were lysed in lysis buffer [0.5% Nonidet P-40 (IGEPAL® CA-630 Sigma-Aldrich I3021), 137 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 1.8 mM KH2 PO4 , 4 mM sodium orthovanadate, 20 mM glycerol-2-phosphate, 10 mM sodium fluoride, 1 mM sodium pyrophosphate], together with complete protease and Phospho-STOP inhibitor mixtures (Roche Applied Science, 05 892 970 001) for 30 min on ice and cleared by centrifugation for 10 min at 20,817 g force. .. After the protein concentration had been determined with the Pierce BCA protein assay kit (ThermoFisher Scientific, 23225), cell lysates were boiled for 5 min at 95°C in reducing Laemmli buffer (Life Technologies, NP0007) supplemented with 100 mM 1,4-dithiothreitol (ThermoFisher Scientific, 20290).

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    Thermo Fisher dtt
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    Thermo Fisher 1 4 dithiothreitol
    1 4 Dithiothreitol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1 4 dithiothreitol/product/Thermo Fisher
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