m dtt dithiothreitol  (Millipore)


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  • 99
    Name:
    L Dithiothreitol
    Description:

    Catalog Number:
    d9760
    Price:
    None
    Applications:
    L-DTT [(2R,3R)-1,4-dimercapto-2,3-butanediol; L-dithiothreitol], a chiral bidentate dithiol with two stereogenic centers, may be used in chiroptical response research. L-DDT is proposed as a stereoselective reducing agent for disulfide bridges in complex molecules.
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    Structured Review

    Millipore m dtt dithiothreitol
    L Dithiothreitol

    https://www.bioz.com/result/m dtt dithiothreitol/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    m dtt dithiothreitol - by Bioz Stars, 2020-09
    99/100 stars

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    In Vitro:

    Article Title: Characterization of pertussis-like toxin from Salmonella spp. that catalyzes ADP-ribosylation of G proteins
    Article Snippet: .. The in vitro ADP-ribosyltransferase reaction mixture (20 μl) contained 0.1 M Tris-HCl (pH 7.6), 0.1 mM ATP, 20 mM DTT, 5 mM thymidine, 10 μM β-NAD, and 0.1 μg of G protein from the bovine brain (Calbiochem-Novabiochem) or 5.6 μg of membrane proteins isolated from RAW 264.7 cells, and either the test substance or Ptx (Biomol, Hamburg, Germany). .. Ptx was preactivated by incubation in 50 mM Tris-HCl (pH 7.5) containing 50 mM DTT at 37 °C for 20 min.

    Isolation:

    Article Title: Characterization of pertussis-like toxin from Salmonella spp. that catalyzes ADP-ribosylation of G proteins
    Article Snippet: .. The in vitro ADP-ribosyltransferase reaction mixture (20 μl) contained 0.1 M Tris-HCl (pH 7.6), 0.1 mM ATP, 20 mM DTT, 5 mM thymidine, 10 μM β-NAD, and 0.1 μg of G protein from the bovine brain (Calbiochem-Novabiochem) or 5.6 μg of membrane proteins isolated from RAW 264.7 cells, and either the test substance or Ptx (Biomol, Hamburg, Germany). .. Ptx was preactivated by incubation in 50 mM Tris-HCl (pH 7.5) containing 50 mM DTT at 37 °C for 20 min.

    Nuclear Magnetic Resonance:

    Article Title: Structure of the Parkin in-between-ring domain provides insights for E3-ligase dysfunction in autosomal recessive Parkinson's disease
    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. .. NMR experiments were acquired on a Varian Inova 600 MHz spectrometer using a xyz -gradient triple-resonance probe.

    other:

    Article Title: Synergism between a foldase and an unfoldase: reciprocal dependence between the thioredoxin-like activity of DnaJ and the polypeptide-unfolding activity of DnaK
    Article Snippet: Insulin turbidity assay The insulin turbidity assay was performed as described in Holmgren ( ) with following modifications; Native insulin (0.13 mM) supplemented either with thioredoxin (1 μM and 10 μM), DnaJ (30 μM) or ΔDnaJ (30 μM) in presence of 0.45 mM DTT (Sigma-Aldrich).

    Activity Assay:

    Article Title: Structure of the Neurospora SET Domain Protein DIM-5, a Histone H3 Lysine Methyltransferase
    Article Snippet: .. The activity was assayed in a 20 µl reaction containing 50 mM glycine (pH 9.8), 2 mM DTT, 40–80 µM unlabeled AdoMet (Sigma), 0.5 µCi [methyl-3 H]AdoMet (78 Ci/mmol, NEN NET155H), 0.25–0.5 µg of DIM-5 protein, and 2–5 µg histones (calf thymus histones Sigma H4524, Roche 223565, or recombinant chicken erythrocyte histones, a gift from Dr. V. Ramakrishnan). .. The reaction was incubated at room temperature for 10–15 min and methylation was analyzed either by SDS-PAGE and fluorography or by precipitation with 20% TCA, filtration (Milipore GF/F filter), washing, and liquid scintillation counting.

    Protease Inhibitor:

    Article Title: ER Stress-Induced Clearance of Misfolded GPI-Anchored Proteins via the Secretory Pathway
    Article Snippet: .. Drug Treatments Working concentrations are 0.1 μM thapsigargin, 0.5 mM dithiothreitol, 5 mM methyl-β-cyclodextrin, and 1 μM brefeldin A. Lysosomal protease inhibitors refer to 125 μM leupeptin + protease inhibitor cocktail (Sigma P8340) diluted 1:100. .. Microscopy All fixed and live-cell imaging, excluding FRAP, were performed using the Marianas spinning disk confocal system (Intelligent Imaging Innovations) attached to a Zeiss Observer.Z1 microscope (Carl Zeiss MicroImaging) and analyzed by Slidebook software.

    Recombinant:

    Article Title: Structure of the Neurospora SET Domain Protein DIM-5, a Histone H3 Lysine Methyltransferase
    Article Snippet: .. The activity was assayed in a 20 µl reaction containing 50 mM glycine (pH 9.8), 2 mM DTT, 40–80 µM unlabeled AdoMet (Sigma), 0.5 µCi [methyl-3 H]AdoMet (78 Ci/mmol, NEN NET155H), 0.25–0.5 µg of DIM-5 protein, and 2–5 µg histones (calf thymus histones Sigma H4524, Roche 223565, or recombinant chicken erythrocyte histones, a gift from Dr. V. Ramakrishnan). .. The reaction was incubated at room temperature for 10–15 min and methylation was analyzed either by SDS-PAGE and fluorography or by precipitation with 20% TCA, filtration (Milipore GF/F filter), washing, and liquid scintillation counting.

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  • 89
    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: 89/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ubiquitination buffer/product/Millipore
    Average 89 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
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    80
    Millipore aconitase buffer
    RT-PCR analysis of <t>aconitase</t> gene expression. Levels of NtACO-1 and β-tubulin mRNA were measured by RT-PCR. cDNA prepared from total RNA by RT was amplified and the RT-PCR products electrophoresed in a 1.75% agarose gel. Gels were stained with ethidium bromide and photographed. β-Tubulin was used as a constitutively expressed control for each RT-PCR reaction. Lane 1, Leaf cDNA; lane 2, mature flower cDNA; lane 3, flower bud cDNA; lane 4, petal cDNA; lane 5, anther cDNA; lane 6, ovary cDNA; lane 7, cDNA from leaves treated 16 h with recombinant rat neuronal NOS plus cofactors and substrate; lane 8, cDNA from leaves treated with only NOS cofactors and substrate.
    Aconitase Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore proteasome buffer
    GAr-carrying chimeras are partially processed by the 26 S <t>proteasome.</t> a , H1299 cells expressing p53-GAr were targeted for 26 S-dependent degradation by overexpressing Mdm2. Immunoblotting using an anti-GAr-specific polyclonal serum revealed a band
    Proteasome Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 88 stars, based on 2 article reviews
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    86
    Millipore tubulin polymerization buffer
    CRMP4 promotes microtubule splaying through its <t>tubulin-binding</t> domain. A , hippocampal neurons from littermate control mice (+/+) or from CRMP4−/− mice transduced with an increasing multiplicity of infection ( MOI ) of wild-type CRMP4 virus
    Tubulin Polymerization Buffer, 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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    Image Search Results


    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

    RT-PCR analysis of aconitase gene expression. Levels of NtACO-1 and β-tubulin mRNA were measured by RT-PCR. cDNA prepared from total RNA by RT was amplified and the RT-PCR products electrophoresed in a 1.75% agarose gel. Gels were stained with ethidium bromide and photographed. β-Tubulin was used as a constitutively expressed control for each RT-PCR reaction. Lane 1, Leaf cDNA; lane 2, mature flower cDNA; lane 3, flower bud cDNA; lane 4, petal cDNA; lane 5, anther cDNA; lane 6, ovary cDNA; lane 7, cDNA from leaves treated 16 h with recombinant rat neuronal NOS plus cofactors and substrate; lane 8, cDNA from leaves treated with only NOS cofactors and substrate.

    Journal: Plant Physiology

    Article Title: Nitric Oxide Modulates the Activity of Tobacco Aconitase 1

    doi:

    Figure Lengend Snippet: RT-PCR analysis of aconitase gene expression. Levels of NtACO-1 and β-tubulin mRNA were measured by RT-PCR. cDNA prepared from total RNA by RT was amplified and the RT-PCR products electrophoresed in a 1.75% agarose gel. Gels were stained with ethidium bromide and photographed. β-Tubulin was used as a constitutively expressed control for each RT-PCR reaction. Lane 1, Leaf cDNA; lane 2, mature flower cDNA; lane 3, flower bud cDNA; lane 4, petal cDNA; lane 5, anther cDNA; lane 6, ovary cDNA; lane 7, cDNA from leaves treated 16 h with recombinant rat neuronal NOS plus cofactors and substrate; lane 8, cDNA from leaves treated with only NOS cofactors and substrate.

    Article Snippet: Extracts were desalted on a PD-10 column (Pharmacia Biotech, Piscataway, NJ) equilibrated with aconitase buffer (20 m m imidazole, pH 7.5, 2 m m citrate, 1 m m DTT, 1 m m EDTA, and 10% [v/v] glycerol) and concentrated in a Centricon-10 column (Millipore, Bedford, MA).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Amplification, Agarose Gel Electrophoresis, Staining, Recombinant

    SDS-PAGE analysis of tobacco aconitase purification. An aliquot of protein sample from each purification step was analyzed by 8% acrylamide SDS-PAGE. The Coomassie blue-stained gel is shown. Lane 1, 10-kD protein ladder; lane 2, total extract; lane 3, 45% to 75% (NH 4 ) 2 SO 4 fraction; lane 4, pooled fractions from the Sephadex G100 column; lane 5, pooled fractions from the phenyl- sepharose column; lane 6, pooled fractions from the Q-sepharose column; lane 7, pooled fractions from the Mono-Q column; lane 8, pooled fractions from the Sephadex 200 HR column. The 50- and 110-kD proteins of a 10-kD protein marker ladder are indicated on the left. The position of the putative aconitase is denoted by an arrow at the right.

    Journal: Plant Physiology

    Article Title: Nitric Oxide Modulates the Activity of Tobacco Aconitase 1

    doi:

    Figure Lengend Snippet: SDS-PAGE analysis of tobacco aconitase purification. An aliquot of protein sample from each purification step was analyzed by 8% acrylamide SDS-PAGE. The Coomassie blue-stained gel is shown. Lane 1, 10-kD protein ladder; lane 2, total extract; lane 3, 45% to 75% (NH 4 ) 2 SO 4 fraction; lane 4, pooled fractions from the Sephadex G100 column; lane 5, pooled fractions from the phenyl- sepharose column; lane 6, pooled fractions from the Q-sepharose column; lane 7, pooled fractions from the Mono-Q column; lane 8, pooled fractions from the Sephadex 200 HR column. The 50- and 110-kD proteins of a 10-kD protein marker ladder are indicated on the left. The position of the putative aconitase is denoted by an arrow at the right.

    Article Snippet: Extracts were desalted on a PD-10 column (Pharmacia Biotech, Piscataway, NJ) equilibrated with aconitase buffer (20 m m imidazole, pH 7.5, 2 m m citrate, 1 m m DTT, 1 m m EDTA, and 10% [v/v] glycerol) and concentrated in a Centricon-10 column (Millipore, Bedford, MA).

    Techniques: SDS Page, Purification, Staining, Marker

    GAr-carrying chimeras are partially processed by the 26 S proteasome. a , H1299 cells expressing p53-GAr were targeted for 26 S-dependent degradation by overexpressing Mdm2. Immunoblotting using an anti-GAr-specific polyclonal serum revealed a band

    Journal:

    Article Title: Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing *Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing * S⃞

    doi: 10.1074/jbc.M803290200

    Figure Lengend Snippet: GAr-carrying chimeras are partially processed by the 26 S proteasome. a , H1299 cells expressing p53-GAr were targeted for 26 S-dependent degradation by overexpressing Mdm2. Immunoblotting using an anti-GAr-specific polyclonal serum revealed a band

    Article Snippet: The beads were washed with phosphate-buffered saline and lysis buffer and then incubated in 26 S proteasome buffer (20 m m Tris-HCl, pH 7.5, 20 m m NaCl, 10 m m MgCl2 , 0.25 m m ATP, 1 m m dithiothreitol) with 400 ng of bacterially expressed hMdm2, 50 ng of E1 (Calbiochem), 50 ng of UbcH5 (Calbiochem), and 1 μg of purified 26 S proteasome (BioMol) at 37 °C for 3 h. The reaction was terminated by adding SDS loading buffer and boiling for 15 min at 85 °C.

    Techniques: Expressing

    Model for GAr-dependent interference with the 26 S proteasome. Fusion of GAr far from the natural unfolding tag of a protein ( e.g. in the N terminus of p53) does not prevent 26 S-dependent degradation and results in the complete degradation of the

    Journal:

    Article Title: Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing *Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing * S⃞

    doi: 10.1074/jbc.M803290200

    Figure Lengend Snippet: Model for GAr-dependent interference with the 26 S proteasome. Fusion of GAr far from the natural unfolding tag of a protein ( e.g. in the N terminus of p53) does not prevent 26 S-dependent degradation and results in the complete degradation of the

    Article Snippet: The beads were washed with phosphate-buffered saline and lysis buffer and then incubated in 26 S proteasome buffer (20 m m Tris-HCl, pH 7.5, 20 m m NaCl, 10 m m MgCl2 , 0.25 m m ATP, 1 m m dithiothreitol) with 400 ng of bacterially expressed hMdm2, 50 ng of E1 (Calbiochem), 50 ng of UbcH5 (Calbiochem), and 1 μg of purified 26 S proteasome (BioMol) at 37 °C for 3 h. The reaction was terminated by adding SDS loading buffer and boiling for 15 min at 85 °C.

    Techniques:

    The 26 S proteasome degrades the N-terminal part of GAr-carrying chimeras. a , fusion of p53 to either end of GAr (HA-p53-GAr-p53) results in degradation of the p53 sequence linked to the N terminus of GAr and the release of the free GAr-p53 * . Treatment

    Journal:

    Article Title: Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing *Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing * S⃞

    doi: 10.1074/jbc.M803290200

    Figure Lengend Snippet: The 26 S proteasome degrades the N-terminal part of GAr-carrying chimeras. a , fusion of p53 to either end of GAr (HA-p53-GAr-p53) results in degradation of the p53 sequence linked to the N terminus of GAr and the release of the free GAr-p53 * . Treatment

    Article Snippet: The beads were washed with phosphate-buffered saline and lysis buffer and then incubated in 26 S proteasome buffer (20 m m Tris-HCl, pH 7.5, 20 m m NaCl, 10 m m MgCl2 , 0.25 m m ATP, 1 m m dithiothreitol) with 400 ng of bacterially expressed hMdm2, 50 ng of E1 (Calbiochem), 50 ng of UbcH5 (Calbiochem), and 1 μg of purified 26 S proteasome (BioMol) at 37 °C for 3 h. The reaction was terminated by adding SDS loading buffer and boiling for 15 min at 85 °C.

    Techniques: Sequencing

    The GAr is a specific regulator of the 26 S proteasome. a , an 80-amino acid-long poly(Q) repeat sequence was fused to p53 (p53-poly(Q) and poly(Q)-p53) and expressed in H1299 cells in the presence or absence of Mdm2. The specific proteasome inhibitor

    Journal:

    Article Title: Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing *Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing * S⃞

    doi: 10.1074/jbc.M803290200

    Figure Lengend Snippet: The GAr is a specific regulator of the 26 S proteasome. a , an 80-amino acid-long poly(Q) repeat sequence was fused to p53 (p53-poly(Q) and poly(Q)-p53) and expressed in H1299 cells in the presence or absence of Mdm2. The specific proteasome inhibitor

    Article Snippet: The beads were washed with phosphate-buffered saline and lysis buffer and then incubated in 26 S proteasome buffer (20 m m Tris-HCl, pH 7.5, 20 m m NaCl, 10 m m MgCl2 , 0.25 m m ATP, 1 m m dithiothreitol) with 400 ng of bacterially expressed hMdm2, 50 ng of E1 (Calbiochem), 50 ng of UbcH5 (Calbiochem), and 1 μg of purified 26 S proteasome (BioMol) at 37 °C for 3 h. The reaction was terminated by adding SDS loading buffer and boiling for 15 min at 85 °C.

    Techniques: Sequencing

    Partial processing of GAr-carrying chimeras by the 26 S proteasome is preceded by an endoproteolytic cleavage. GAr was fused to either end of p53 (GAr-p53-GAr-HA) to protect the substrate from end first-mediated degradation by the proteasome. a , GAr-p53-GAr-HA

    Journal:

    Article Title: Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing *Gly-Ala Repeats Induce Position- and Substrate-specific Regulation of 26 S Proteasome-dependent Partial Processing * S⃞

    doi: 10.1074/jbc.M803290200

    Figure Lengend Snippet: Partial processing of GAr-carrying chimeras by the 26 S proteasome is preceded by an endoproteolytic cleavage. GAr was fused to either end of p53 (GAr-p53-GAr-HA) to protect the substrate from end first-mediated degradation by the proteasome. a , GAr-p53-GAr-HA

    Article Snippet: The beads were washed with phosphate-buffered saline and lysis buffer and then incubated in 26 S proteasome buffer (20 m m Tris-HCl, pH 7.5, 20 m m NaCl, 10 m m MgCl2 , 0.25 m m ATP, 1 m m dithiothreitol) with 400 ng of bacterially expressed hMdm2, 50 ng of E1 (Calbiochem), 50 ng of UbcH5 (Calbiochem), and 1 μg of purified 26 S proteasome (BioMol) at 37 °C for 3 h. The reaction was terminated by adding SDS loading buffer and boiling for 15 min at 85 °C.

    Techniques:

    CRMP4 promotes microtubule splaying through its tubulin-binding domain. A , hippocampal neurons from littermate control mice (+/+) or from CRMP4−/− mice transduced with an increasing multiplicity of infection ( MOI ) of wild-type CRMP4 virus

    Journal: The Journal of Biological Chemistry

    Article Title: Collapsin Response Mediator Protein 4 Regulates Growth Cone Dynamics through the Actin and Microtubule Cytoskeleton *

    doi: 10.1074/jbc.M114.570440

    Figure Lengend Snippet: CRMP4 promotes microtubule splaying through its tubulin-binding domain. A , hippocampal neurons from littermate control mice (+/+) or from CRMP4−/− mice transduced with an increasing multiplicity of infection ( MOI ) of wild-type CRMP4 virus

    Article Snippet: Beads were then washed five times with 10 bead volumes of washing buffer (50 m m Tris-HCl, pH 7.5, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) and eluted with 20 m m glutathione in elution buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) followed by concentration and buffer exchange to tubulin polymerization buffer (10 m m Tris-HCl, pH 7.5, 2 m m MgCl2 , 1 m m DTT) with an Amicon Ultra-4 30,000 Centricon column concentrator (Millipore).

    Techniques: Binding Assay, Mouse Assay, Transduction, Infection

    CRMP4 regulates axon outgrowth and growth cone size. A , hippocampal neurons from CRMP4+/+ and CRMP4−/− mice were cultured for 3 DIV and stained with anti-βIII-tubulin antibody. Scale bar , 25 μm. B and C , the average axon

    Journal: The Journal of Biological Chemistry

    Article Title: Collapsin Response Mediator Protein 4 Regulates Growth Cone Dynamics through the Actin and Microtubule Cytoskeleton *

    doi: 10.1074/jbc.M114.570440

    Figure Lengend Snippet: CRMP4 regulates axon outgrowth and growth cone size. A , hippocampal neurons from CRMP4+/+ and CRMP4−/− mice were cultured for 3 DIV and stained with anti-βIII-tubulin antibody. Scale bar , 25 μm. B and C , the average axon

    Article Snippet: Beads were then washed five times with 10 bead volumes of washing buffer (50 m m Tris-HCl, pH 7.5, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) and eluted with 20 m m glutathione in elution buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) followed by concentration and buffer exchange to tubulin polymerization buffer (10 m m Tris-HCl, pH 7.5, 2 m m MgCl2 , 1 m m DTT) with an Amicon Ultra-4 30,000 Centricon column concentrator (Millipore).

    Techniques: Mouse Assay, Cell Culture, Staining

    CRMP4 activity toward microtubules promotes growth cone expansion and axon outgrowth. A and C , hippocampal neurons from CRMP4−/− mice transduced with GFP or CRMP4 rescue constructs and stained with anti-βIII-tubulin ( A ) or anti-βIII-tubulin

    Journal: The Journal of Biological Chemistry

    Article Title: Collapsin Response Mediator Protein 4 Regulates Growth Cone Dynamics through the Actin and Microtubule Cytoskeleton *

    doi: 10.1074/jbc.M114.570440

    Figure Lengend Snippet: CRMP4 activity toward microtubules promotes growth cone expansion and axon outgrowth. A and C , hippocampal neurons from CRMP4−/− mice transduced with GFP or CRMP4 rescue constructs and stained with anti-βIII-tubulin ( A ) or anti-βIII-tubulin

    Article Snippet: Beads were then washed five times with 10 bead volumes of washing buffer (50 m m Tris-HCl, pH 7.5, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) and eluted with 20 m m glutathione in elution buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) followed by concentration and buffer exchange to tubulin polymerization buffer (10 m m Tris-HCl, pH 7.5, 2 m m MgCl2 , 1 m m DTT) with an Amicon Ultra-4 30,000 Centricon column concentrator (Millipore).

    Techniques: Activity Assay, Mouse Assay, Transduction, Construct, Staining

    CRMP4 expression in hippocampal neurons. A , Western blot of an E18 wild-type hippocampal lysate probed with anti-CRMP4 antibody. B , 3 DIV wild-type hippocampal neurons stained with rhodamine phalloidin and anti-βIII-tubulin and anti-CRMP4 antibodies.

    Journal: The Journal of Biological Chemistry

    Article Title: Collapsin Response Mediator Protein 4 Regulates Growth Cone Dynamics through the Actin and Microtubule Cytoskeleton *

    doi: 10.1074/jbc.M114.570440

    Figure Lengend Snippet: CRMP4 expression in hippocampal neurons. A , Western blot of an E18 wild-type hippocampal lysate probed with anti-CRMP4 antibody. B , 3 DIV wild-type hippocampal neurons stained with rhodamine phalloidin and anti-βIII-tubulin and anti-CRMP4 antibodies.

    Article Snippet: Beads were then washed five times with 10 bead volumes of washing buffer (50 m m Tris-HCl, pH 7.5, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) and eluted with 20 m m glutathione in elution buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 5 m m MgCl2 , 1 m m DTT) followed by concentration and buffer exchange to tubulin polymerization buffer (10 m m Tris-HCl, pH 7.5, 2 m m MgCl2 , 1 m m DTT) with an Amicon Ultra-4 30,000 Centricon column concentrator (Millipore).

    Techniques: Expressing, Western Blot, Staining