pkcβii fragments  (Roche)


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    Structured Review

    Roche pkcβii fragments
    <t>PKCβII</t> is required for the responsiveness of DRG neurons to heat and capsaicin. A , Representative inward currents from a DRG neuron under control conditions (left), from a neuron transfected with GFP-PKCβII (middle), and from a neuron
    Pkcβii Fragments, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 30099 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pkcβii fragments/product/Roche
    Average 86 stars, based on 30099 article reviews
    Price from $9.99 to $1999.99
    pkcβii fragments - by Bioz Stars, 2020-09
    86/100 stars

    Images

    1) Product Images from "The Basal Thermal Sensitivity of the TRPV1 Ion Channel Is Determined by PKCβII"

    Article Title: The Basal Thermal Sensitivity of the TRPV1 Ion Channel Is Determined by PKCβII

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0278-14.2014

    PKCβII is required for the responsiveness of DRG neurons to heat and capsaicin. A , Representative inward currents from a DRG neuron under control conditions (left), from a neuron transfected with GFP-PKCβII (middle), and from a neuron
    Figure Legend Snippet: PKCβII is required for the responsiveness of DRG neurons to heat and capsaicin. A , Representative inward currents from a DRG neuron under control conditions (left), from a neuron transfected with GFP-PKCβII (middle), and from a neuron

    Techniques Used: Transfection

    TRPV1 causes the translocation of PKCβII in DRG neurons. A , Example of double immunostaining of PKCε and TRPV1 in DRG neurons. PKCε was expressed in the cytoplasm in all DRG neurons regardless of whether TRPV1 was coexpressed.
    Figure Legend Snippet: TRPV1 causes the translocation of PKCβII in DRG neurons. A , Example of double immunostaining of PKCε and TRPV1 in DRG neurons. PKCε was expressed in the cytoplasm in all DRG neurons regardless of whether TRPV1 was coexpressed.

    Techniques Used: Translocation Assay, Double Immunostaining

    Downregulation of TRPV1 by PKCβII depends on the phosphorylation of TRPV1 at Thr705. A , B , Catalytic activity of PKCβII is essential for the downregulation of TRPV1. Tet-On HEK-293 cells were transfected with TRPV1–V5 together
    Figure Legend Snippet: Downregulation of TRPV1 by PKCβII depends on the phosphorylation of TRPV1 at Thr705. A , B , Catalytic activity of PKCβII is essential for the downregulation of TRPV1. Tet-On HEK-293 cells were transfected with TRPV1–V5 together

    Techniques Used: Activity Assay, Transfection

    PKCβII increases the sensitivity of TRPV1 through a phosphorylation-dependent mechanism. A , Knock down of PKCβII and PKCε by shRNAs. HEK-293 cells were transfected with scrambled or PKCβII shRNA (top two blots) or PKCε
    Figure Legend Snippet: PKCβII increases the sensitivity of TRPV1 through a phosphorylation-dependent mechanism. A , Knock down of PKCβII and PKCε by shRNAs. HEK-293 cells were transfected with scrambled or PKCβII shRNA (top two blots) or PKCε

    Techniques Used: Transfection, shRNA

    Downregulation of TRPV1 and PKCβII. A , Expression of TRPV1 was downregulated by PKCα and PKCβII, but not by PKCδ or PKCε. HEK 293 cells were transfected with TRPV1–V5 together with different PKC isoforms
    Figure Legend Snippet: Downregulation of TRPV1 and PKCβII. A , Expression of TRPV1 was downregulated by PKCα and PKCβII, but not by PKCδ or PKCε. HEK 293 cells were transfected with TRPV1–V5 together with different PKC isoforms

    Techniques Used: Expressing, Transfection

    TRPV1 binds directly to PKCβII. A , Localization of PKCβII (left) and TRPV1 (middle) in HEK 293 cells expressing GFP-PKCβII and TRPV1. On the right is a merged image. Scale bars, 10 μm. B , PKCβII was coprecipitated
    Figure Legend Snippet: TRPV1 binds directly to PKCβII. A , Localization of PKCβII (left) and TRPV1 (middle) in HEK 293 cells expressing GFP-PKCβII and TRPV1. On the right is a merged image. Scale bars, 10 μm. B , PKCβII was coprecipitated

    Techniques Used: Expressing

    Heat sensitivity of TRPV1 is determined by the phosphorylation of T705 induced by PKCβII. A , Example of inward current (top left trace) induced by a temperature ramp shown under Tet-On HEK-293 cells expressing T705A TRPV1. Right, current elicited
    Figure Legend Snippet: Heat sensitivity of TRPV1 is determined by the phosphorylation of T705 induced by PKCβII. A , Example of inward current (top left trace) induced by a temperature ramp shown under Tet-On HEK-293 cells expressing T705A TRPV1. Right, current elicited

    Techniques Used: Expressing

    PKCε, but not PKCβII, is involved in the potentiation of TRPV1. A – C , Examples of whole-cell inward currents activated by capsaicin (50 n m , 5 s) from a HEK 293 cell expressing TRPV1 and the bradykinin B2 receptor ( A ; control) or
    Figure Legend Snippet: PKCε, but not PKCβII, is involved in the potentiation of TRPV1. A – C , Examples of whole-cell inward currents activated by capsaicin (50 n m , 5 s) from a HEK 293 cell expressing TRPV1 and the bradykinin B2 receptor ( A ; control) or

    Techniques Used: Expressing

    Heat sensitivity of TRPV1 is increased by PKCβII. A , Representative inward currents carried by TRPV1 (top left trace) induced by a temperature ramp shown underneath in a Tet-On HEK-293 cell expressing TRPV1 and PKCβII-pTRE2. Right, Inward
    Figure Legend Snippet: Heat sensitivity of TRPV1 is increased by PKCβII. A , Representative inward currents carried by TRPV1 (top left trace) induced by a temperature ramp shown underneath in a Tet-On HEK-293 cell expressing TRPV1 and PKCβII-pTRE2. Right, Inward

    Techniques Used: Expressing

    Related Articles

    Pull Down Assay:

    Article Title: Positive feedback regulation of p53 transactivity by DNA damage-induced ISG15 modification
    Article Snippet: .. Immunoprecipitation and pull-down assay For immunoprecipitation, cell lysates were prepared in buffer-A consisting of 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1 mM NEM, 1 mM sodium orthovanadate, 1 mM NaF, 1 mM PMSF, 0.2% (v/v) Triton X-100 and 1X protease inhibitor cocktail (Roche). .. The cell lysates were incubated with appropriate antibodies for 1 h at 4 °C and then with protein A-conjugated Sepharose for the next 1 h. For Ni2+ -NTA agarose (NTA: Qiagen) pull-down assay, the cell lysates were prepared in buffer-A containing 10 mM imidazole.

    Protease Inhibitor:

    Article Title: A Novel Cell Lysis Approach Reveals That Caspase-2 Rapidly Translocates from the Nucleus to the Cytoplasm in Response to Apoptotic Stimuli
    Article Snippet: .. The cells were then washed with ice-cold isotonic saline and lysed in ice-cold lysis buffer (50 mM Tris-HCl (pH 7.4), 150 mM KCl, 0.5% Triton X-100 with Roche mini-complete protease inhibitor) by repeated vortexing at maximal setting and keeping the cells on ice for 10 min. Lysing cells with Triton X-100 at isotonic salt concentrations has been shown to be a rapid and effective way to isolate purified nuclei . .. The lysates were centrifuged for 10 min at 5,000 g, the extra-nuclear faction was transferred to a new tube and the nuclear pellets were washed with lysis buffer.

    Article Title: Positive feedback regulation of p53 transactivity by DNA damage-induced ISG15 modification
    Article Snippet: .. Immunoprecipitation and pull-down assay For immunoprecipitation, cell lysates were prepared in buffer-A consisting of 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1 mM NEM, 1 mM sodium orthovanadate, 1 mM NaF, 1 mM PMSF, 0.2% (v/v) Triton X-100 and 1X protease inhibitor cocktail (Roche). .. The cell lysates were incubated with appropriate antibodies for 1 h at 4 °C and then with protein A-conjugated Sepharose for the next 1 h. For Ni2+ -NTA agarose (NTA: Qiagen) pull-down assay, the cell lysates were prepared in buffer-A containing 10 mM imidazole.

    Article Title: SRPK1 acetylation modulates alternative splicing to regulate cisplatin resistance in breast cancer cells
    Article Snippet: .. Immunoprecipitation and co-immunoprecipitation The cells were lysed in a lysis buffer (50 mM pH 8.0 Tris-HCl, 150 mM NaCl, 1% NP-40, 0.05% sodium deoxycholate) supplemented with cOmpleteTM Protease Inhibitor (Roche) for 30 min at 4 °C. .. The supernatant was collected after centrifugation and precleared with Protein G Plus slurry (Santa Cruz).

    Generated:

    Article Title: A versatile system to introduce clusters of genomic double-strand breaks in large cell populations
    Article Snippet: .. Immunoprecipitation and Immunoblotting Cell lysates were generated using RIPA buffer (10 mM Tris-HCl at ph 8.0, 1 mM EDTA, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, 150 mM NaCl) containing protease inhibitors (Roche complete) and 100U/ml benzonase (Merck). .. Immunoprecipitation (IP) was performed with protein A-coupled magnetic beads (Invitrogen Dynal).

    Purification:

    Article Title: A Novel Cell Lysis Approach Reveals That Caspase-2 Rapidly Translocates from the Nucleus to the Cytoplasm in Response to Apoptotic Stimuli
    Article Snippet: .. The cells were then washed with ice-cold isotonic saline and lysed in ice-cold lysis buffer (50 mM Tris-HCl (pH 7.4), 150 mM KCl, 0.5% Triton X-100 with Roche mini-complete protease inhibitor) by repeated vortexing at maximal setting and keeping the cells on ice for 10 min. Lysing cells with Triton X-100 at isotonic salt concentrations has been shown to be a rapid and effective way to isolate purified nuclei . .. The lysates were centrifuged for 10 min at 5,000 g, the extra-nuclear faction was transferred to a new tube and the nuclear pellets were washed with lysis buffer.

    Real-time Polymerase Chain Reaction:

    Article Title: Maintaining mRNA Integrity during Decalcification of Mineralized Tissues
    Article Snippet: .. Quantitative PCR RNA isolated from EDTA or RNAlater /EDTA decalcified whole tibiae (500 ng total RNA) was reverse-transcribed as per the manufacturer’s protocol in a 20µL reaction using the Transcriptor High Fidelity cDNA Synthesis Kit (Roche, Germany). .. Quantitative PCR (qPCR) reactions containing 2.5 ng cDNA were prepared to 5µL in 384 well PCR plates (Roche) using the LightCycler® 480 SYBR Green I Master (Roche) as per the manufacturer’s instructions and run on a LC480 II thermo cycler (Roche).

    Immunoprecipitation:

    Article Title: A versatile system to introduce clusters of genomic double-strand breaks in large cell populations
    Article Snippet: .. Immunoprecipitation and Immunoblotting Cell lysates were generated using RIPA buffer (10 mM Tris-HCl at ph 8.0, 1 mM EDTA, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, 150 mM NaCl) containing protease inhibitors (Roche complete) and 100U/ml benzonase (Merck). .. Immunoprecipitation (IP) was performed with protein A-coupled magnetic beads (Invitrogen Dynal).

    Article Title: Positive feedback regulation of p53 transactivity by DNA damage-induced ISG15 modification
    Article Snippet: .. Immunoprecipitation and pull-down assay For immunoprecipitation, cell lysates were prepared in buffer-A consisting of 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1 mM NEM, 1 mM sodium orthovanadate, 1 mM NaF, 1 mM PMSF, 0.2% (v/v) Triton X-100 and 1X protease inhibitor cocktail (Roche). .. The cell lysates were incubated with appropriate antibodies for 1 h at 4 °C and then with protein A-conjugated Sepharose for the next 1 h. For Ni2+ -NTA agarose (NTA: Qiagen) pull-down assay, the cell lysates were prepared in buffer-A containing 10 mM imidazole.

    Article Title: SRPK1 acetylation modulates alternative splicing to regulate cisplatin resistance in breast cancer cells
    Article Snippet: .. Immunoprecipitation and co-immunoprecipitation The cells were lysed in a lysis buffer (50 mM pH 8.0 Tris-HCl, 150 mM NaCl, 1% NP-40, 0.05% sodium deoxycholate) supplemented with cOmpleteTM Protease Inhibitor (Roche) for 30 min at 4 °C. .. The supernatant was collected after centrifugation and precleared with Protein G Plus slurry (Santa Cruz).

    Incubation:

    Article Title: The lipid phosphatase Synaptojanin 1 undergoes a significant alteration in expression and solubility and is associated with brain lesions in Alzheimer’s disease
    Article Snippet: .. Preparation of brain homogenates for biochemical analysisAbout 200 mg of frozen T1 isocortex was homogenised as reported [ , ] in 10 volumes of ice-cold RIPA buffer containing 50 mM Tris pH 7.4 containing 150 mM NaCl, 1% NP-40, 0.25% sodium deoxycholate, 5 mM EDTA, 1 mM EGTA, Roche complete protease inhibitors, 1 mM PMSF, and phosphatase inhibitor cocktail 2, (Sigma, P-5726) and incubated for 60 min at 4 °C on a rotator. .. One hundred microliter of the total homogenate was supplemented with Laemmli buffer, sonicated on ice and analysed as the total fraction.

    Isolation:

    Article Title: Maintaining mRNA Integrity during Decalcification of Mineralized Tissues
    Article Snippet: .. Quantitative PCR RNA isolated from EDTA or RNAlater /EDTA decalcified whole tibiae (500 ng total RNA) was reverse-transcribed as per the manufacturer’s protocol in a 20µL reaction using the Transcriptor High Fidelity cDNA Synthesis Kit (Roche, Germany). .. Quantitative PCR (qPCR) reactions containing 2.5 ng cDNA were prepared to 5µL in 384 well PCR plates (Roche) using the LightCycler® 480 SYBR Green I Master (Roche) as per the manufacturer’s instructions and run on a LC480 II thermo cycler (Roche).

    Lysis:

    Article Title: A Novel Cell Lysis Approach Reveals That Caspase-2 Rapidly Translocates from the Nucleus to the Cytoplasm in Response to Apoptotic Stimuli
    Article Snippet: .. The cells were then washed with ice-cold isotonic saline and lysed in ice-cold lysis buffer (50 mM Tris-HCl (pH 7.4), 150 mM KCl, 0.5% Triton X-100 with Roche mini-complete protease inhibitor) by repeated vortexing at maximal setting and keeping the cells on ice for 10 min. Lysing cells with Triton X-100 at isotonic salt concentrations has been shown to be a rapid and effective way to isolate purified nuclei . .. The lysates were centrifuged for 10 min at 5,000 g, the extra-nuclear faction was transferred to a new tube and the nuclear pellets were washed with lysis buffer.

    Article Title: SRPK1 acetylation modulates alternative splicing to regulate cisplatin resistance in breast cancer cells
    Article Snippet: .. Immunoprecipitation and co-immunoprecipitation The cells were lysed in a lysis buffer (50 mM pH 8.0 Tris-HCl, 150 mM NaCl, 1% NP-40, 0.05% sodium deoxycholate) supplemented with cOmpleteTM Protease Inhibitor (Roche) for 30 min at 4 °C. .. The supernatant was collected after centrifugation and precleared with Protein G Plus slurry (Santa Cruz).

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