shrimp alkaline phosphatase  (TaKaRa)


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  • 94
    Name:
    Alkaline Phosphatase
    Description:

    Catalog Number:
    2660A
    Price:
    None
    Category:
    Molecular Biology
    Size:
    300 Units
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    Structured Review

    TaKaRa shrimp alkaline phosphatase

    https://www.bioz.com/result/shrimp alkaline phosphatase/product/TaKaRa
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    shrimp alkaline phosphatase - by Bioz Stars, 2021-05
    94/100 stars

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    Incubation:

    Article Title: Serotyping of Streptococcus pneumoniae Based on Capsular Genes Polymorphisms
    Article Snippet: The PCR program consisted in the following steps: 60 s at 94°C followed by 39 cycles of 30 s at 94°C, 30 s at 55°C and 90 s at 72°C. .. Finally, the reaction was incubated at 72°C for 3 min. Then, 3 units of shrimp alkaline phosphatase (Clontech, Mountain View, CA), 7.5 units of exonuclease (Clontech) and 0.25 µl of 50X titanium DNA polymerase (Clontech) were added to the solution, which was incubated at 37°C for 20 min and at 94°C for 10 min. .. This step allows for the degradation of remaining dNTPs and PCR primers that were not used in the multiplex PCR.

    Article Title: Polymorphism study of nine SNPs associated with subjective response to alcohol in Chinese Han, Hui, Tibetan, Mongolian and Uygur populations
    Article Snippet: Multiplex amplification reactions were performed in a ProFlex™ 96-well PCR System (Thermo Fisher Scientific, USA). .. To remove excess primers and deoxy-ribonucleoside triphosphates (dNTPs), the PCR products were treated with 2.5 U of shrimp alkaline phosphatase (SAP) (TaKaRa Biotech, China) and 6 U of exonuclease I (TaKaRa Biotech, China) by incubation at 37 °C for 1 h, followed by enzyme inactivation at 80 °C for 10 min. .. Multiplex SBE reactions and purification of SBE products The extension reaction was performed in a 5 μL final volume containing 1.5 μL of PCR product, 0.15 μL of each extension primer (10 μmol/L) and 1.5 μL of SNaPshot reaction mix (Applied Biosystems, USA).

    Article Title: Metagenomic Analysis of Flaviviridae in Mosquito Viromes Isolated From Yunnan Province in China Reveals Genes From Dengue and Zika Viruses
    Article Snippet: To synthesize dscDNA, anchored random primers were added and incubated 75°C for 5 min and then on ice for 5 min for denaturation. .. Then, 1 μL of Klenow fragment (TaKaRa), 1 μL of 10 mM dNTPs (TaKaRa), 2 μL of 10 × Klenow buffer (TaKaRa), and 6 μL of dd H2 O (TaKaRa) were added and the samples were incubated at 37°C for 60 min, followed by 75°C for 10 min. Then, 0.5 μL of exonuclease I (TaKaRa), 1 μL of shrimp alkaline phosphatase (SAP, TaKaRa, Dalian, China), 5 μL of 10 × phosphatase buffer (TaKaRa), and 24 μL of DEPC H2 O (TaKaRa) were added and incubated at 37°C for 60 min followed by 75°C for 10 min to eliminate the phosphates and the free single-strand nucleic acid in the dscDNA reaction. .. Sequence-independent single-primer amplification (SISPA) and purification of PCR products To increase the quantity of the viral nucleic acids, SISPA was applied to amplify the dscDNA.

    Article Title: N-Terminal Phosphorylation of HP1? Promotes Its Chromatin Binding ▿
    Article Snippet: Mouse and human cell lines were lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl [pH 8.0], 150 mM NaCl, 1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate [SDS]) supplemented with protease inhibitor cocktail (Complete [EDTA-free]; Roche) and 10 mM sodium fluoride (NaF). .. For dephosphorylation, an aliquot of whole-cell lysate (WCL) was incubated with 0.08 U/ml shrimp alkaline phosphatase (SAP) (TaKaRa) for 3 to 6 h at 37°C. .. Western blotting was performed according to a standard protocol.

    Plasmid Preparation:

    Article Title: Intracellular β-Glucosidases CEL1a and CEL1b Are Essential for Cellulase Induction on Lactose in Trichoderma reesei
    Article Snippet: In order to complement the Δ cel1a Δ cel1b strain with CEL1a and CEL1a (E367A) under the control of the native cel1a promoter, the cel1a disruption vector pUC cel1apyr4 ( ) was digested with XbaI and SalI to remove the pyr4 gene, followed by ligation with the hygromycin resistance cassette digested with the same enzymes to create pUC cel1ahph . .. This plasmid was digested with XbaI, dephosphorylated using shrimp alkaline phosphatase (TaKaRa), and further ligated with the coding region for CEL1a to obtain pNE cel1a . .. A mutant cel1a gene encoding a E367A substitution in wild-type (WT) CEL1a was created by oligonucleotide-mediated mutagenesis of the cel1a gene, using a two-step fusion PCR with pNE cel1a as the template.

    Polymerase Chain Reaction:

    Article Title: Putative Precursor Cancer Cells in Human Colorectal Cancer Tissue
    Article Snippet: .. PCR fragments for primer extension were prepared by incubating 7.5 μl PCR product with 0.5 U shrimp alkaline phosphatase (TaKaRa) and 1 U exonuclease I (TaKaRa) for 40 min at 37°C in a final volume of 10 μl, followed by the inactivation of the enzymes for 20 min at 80°C. .. Primer extension was carried out in 5 μl containing 2 μl of treated PCR product, 2.5 μl ABI Prism SNaPshot Multiplex kit (Applied Biosystems) and 0.5 μl extension primers mix (0.2μM each primer).

    Article Title: Mutation screening of the UBE3A gene in Chinese Han population with autism
    Article Snippet: PCR condition contained initial denaturation at 95 °C for 5 min, 30 cycles of 95 °C for 30 s and 62 °C–65 °C for 30 s and 72 °C for 45 s, then 72 °C for 5 min as final extension was performed. .. After purification of PCR products using TaKaRa® Shrimp Alkaline Phosphatase (SAP) and ExonucleaseI (ExonI), sequencing was performed by BigDye Terminator Kit (Applied Biosystems, USA) according to the method by Sanger F, et al. [ ]. .. The sequence information were read using Applied Biosystems (ABI) 3130 Genetic Analyzer (Applied Biosystems, USA).

    Article Title: Two Distinct mtDNA Lineages among Captive African Penguins in Japan
    Article Snippet: PCR products were checked by electrophoresis on 1.5% agarose gel with TAE buffer, and the gel was stained with ethidium bromide. .. PCR products were purified by exonuclease I (Wako, Osaka, Japan) and shrimp alkaline phosphatase (TaKaRa Bio) and were sequenced directly using an ABI PRISM 3130 Genetic Analyzer and BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster, CA, U.S.A.). ..

    Article Title: Polymorphism study of nine SNPs associated with subjective response to alcohol in Chinese Han, Hui, Tibetan, Mongolian and Uygur populations
    Article Snippet: Multiplex amplification reactions were performed in a ProFlex™ 96-well PCR System (Thermo Fisher Scientific, USA). .. To remove excess primers and deoxy-ribonucleoside triphosphates (dNTPs), the PCR products were treated with 2.5 U of shrimp alkaline phosphatase (SAP) (TaKaRa Biotech, China) and 6 U of exonuclease I (TaKaRa Biotech, China) by incubation at 37 °C for 1 h, followed by enzyme inactivation at 80 °C for 10 min. .. Multiplex SBE reactions and purification of SBE products The extension reaction was performed in a 5 μL final volume containing 1.5 μL of PCR product, 0.15 μL of each extension primer (10 μmol/L) and 1.5 μL of SNaPshot reaction mix (Applied Biosystems, USA).

    Purification:

    Article Title: Mutation screening of the UBE3A gene in Chinese Han population with autism
    Article Snippet: PCR condition contained initial denaturation at 95 °C for 5 min, 30 cycles of 95 °C for 30 s and 62 °C–65 °C for 30 s and 72 °C for 45 s, then 72 °C for 5 min as final extension was performed. .. After purification of PCR products using TaKaRa® Shrimp Alkaline Phosphatase (SAP) and ExonucleaseI (ExonI), sequencing was performed by BigDye Terminator Kit (Applied Biosystems, USA) according to the method by Sanger F, et al. [ ]. .. The sequence information were read using Applied Biosystems (ABI) 3130 Genetic Analyzer (Applied Biosystems, USA).

    Article Title: Two Distinct mtDNA Lineages among Captive African Penguins in Japan
    Article Snippet: PCR products were checked by electrophoresis on 1.5% agarose gel with TAE buffer, and the gel was stained with ethidium bromide. .. PCR products were purified by exonuclease I (Wako, Osaka, Japan) and shrimp alkaline phosphatase (TaKaRa Bio) and were sequenced directly using an ABI PRISM 3130 Genetic Analyzer and BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster, CA, U.S.A.). ..

    Sequencing:

    Article Title: Mutation screening of the UBE3A gene in Chinese Han population with autism
    Article Snippet: PCR condition contained initial denaturation at 95 °C for 5 min, 30 cycles of 95 °C for 30 s and 62 °C–65 °C for 30 s and 72 °C for 45 s, then 72 °C for 5 min as final extension was performed. .. After purification of PCR products using TaKaRa® Shrimp Alkaline Phosphatase (SAP) and ExonucleaseI (ExonI), sequencing was performed by BigDye Terminator Kit (Applied Biosystems, USA) according to the method by Sanger F, et al. [ ]. .. The sequence information were read using Applied Biosystems (ABI) 3130 Genetic Analyzer (Applied Biosystems, USA).

    Article Title: Two Distinct mtDNA Lineages among Captive African Penguins in Japan
    Article Snippet: PCR products were checked by electrophoresis on 1.5% agarose gel with TAE buffer, and the gel was stained with ethidium bromide. .. PCR products were purified by exonuclease I (Wako, Osaka, Japan) and shrimp alkaline phosphatase (TaKaRa Bio) and were sequenced directly using an ABI PRISM 3130 Genetic Analyzer and BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster, CA, U.S.A.). ..

    De-Phosphorylation Assay:

    Article Title: N-Terminal Phosphorylation of HP1? Promotes Its Chromatin Binding ▿
    Article Snippet: Mouse and human cell lines were lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl [pH 8.0], 150 mM NaCl, 1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate [SDS]) supplemented with protease inhibitor cocktail (Complete [EDTA-free]; Roche) and 10 mM sodium fluoride (NaF). .. For dephosphorylation, an aliquot of whole-cell lysate (WCL) was incubated with 0.08 U/ml shrimp alkaline phosphatase (SAP) (TaKaRa) for 3 to 6 h at 37°C. .. Western blotting was performed according to a standard protocol.

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  • 94
    TaKaRa shrimp alkaline phosphatase sap
    Identification of phosphorylation sites of mouse HP1α. (A) The phosphorylation patterns of mouse HP1α analyzed by phos-tag Western blotting (phos-tag WB). Whole-cell lysates (WCLs) of NIH 3T3 cells were resolved on an SDS-PAGE gel containing phos-tag acrylamide (phos-tag) or on a standard SDS-PAGE gel (mock). As an unphosphorylated control, <t>SAP-treated</t> <t>WCL</t> was loaded on the same gel. HP1α was detected using an anti-HP1α antibody. (B) Phosphorylation patterns of mouse HP1α in cells arrested at different cell cycle stages. WCLs were prepared from NIH 3T3 cells grown under the indicated conditions and analyzed by phos-tag WB. The arrowhead indicates an HP1α with basal band shift. The asterisk represents an HP1α species with metaphase-associated phosphorylation. The same membrane was immunoblotted with anti-α-tubulin and anti-TIF1β antibodies for loading controls. (C) Amino acid sequence of mouse HP1α. Light- and dark-shaded boxes indicate the CD and CSD, respectively. The sites of serine (S)-to-alanine (A) substitutions introduced in the following experiments are shown in boldface. (D) Phos-tag and standard WB analysis of wild-type and mutant HP1αs. WCLs were prepared from NIH 3T3 cells transiently expressing FLAG-tagged wild-type HP1α or HP1α with N-terminal (N-term: S11-14A) mutations, hinge (Hinge: S92, 93, 95, 97A) mutations, or both (N-term + hinge). Each FLAG-HP1α with an SAP-treated control was analyzed by phos-tag WB using an anti-FLAG antibody. The phosphorylation state of HP1α represented by each protein band was deduced from the distance of migration level of each band (Rf) and is indicated to the right of the phos-tag WB panel. (E) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the N-terminal region. The phosphorylation patterns of the indicated FLAG-HP1α species were analyzed by phos-tag WB using a higher concentration (80 μM) of phos-tag acrylamide. (F) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the hinge region. (G) The numbers of phosphorylated peptides identified by liquid chromatography (LC)-MS/MS analysis of affinity-purified HP1α. (H) Summary of the phosphorylation sites of mouse HP1α deduced from the experiments described above.
    Shrimp Alkaline Phosphatase Sap, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/shrimp alkaline phosphatase sap/product/TaKaRa
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    shrimp alkaline phosphatase sap - by Bioz Stars, 2021-05
    94/100 stars
      Buy from Supplier

    99
    TaKaRa shrimp alkaline phosphatase
    Identification of phosphorylation sites of mouse HP1α. (A) The phosphorylation patterns of mouse HP1α analyzed by phos-tag Western blotting (phos-tag WB). Whole-cell lysates (WCLs) of NIH 3T3 cells were resolved on an SDS-PAGE gel containing phos-tag acrylamide (phos-tag) or on a standard SDS-PAGE gel (mock). As an unphosphorylated control, <t>SAP-treated</t> <t>WCL</t> was loaded on the same gel. HP1α was detected using an anti-HP1α antibody. (B) Phosphorylation patterns of mouse HP1α in cells arrested at different cell cycle stages. WCLs were prepared from NIH 3T3 cells grown under the indicated conditions and analyzed by phos-tag WB. The arrowhead indicates an HP1α with basal band shift. The asterisk represents an HP1α species with metaphase-associated phosphorylation. The same membrane was immunoblotted with anti-α-tubulin and anti-TIF1β antibodies for loading controls. (C) Amino acid sequence of mouse HP1α. Light- and dark-shaded boxes indicate the CD and CSD, respectively. The sites of serine (S)-to-alanine (A) substitutions introduced in the following experiments are shown in boldface. (D) Phos-tag and standard WB analysis of wild-type and mutant HP1αs. WCLs were prepared from NIH 3T3 cells transiently expressing FLAG-tagged wild-type HP1α or HP1α with N-terminal (N-term: S11-14A) mutations, hinge (Hinge: S92, 93, 95, 97A) mutations, or both (N-term + hinge). Each FLAG-HP1α with an SAP-treated control was analyzed by phos-tag WB using an anti-FLAG antibody. The phosphorylation state of HP1α represented by each protein band was deduced from the distance of migration level of each band (Rf) and is indicated to the right of the phos-tag WB panel. (E) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the N-terminal region. The phosphorylation patterns of the indicated FLAG-HP1α species were analyzed by phos-tag WB using a higher concentration (80 μM) of phos-tag acrylamide. (F) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the hinge region. (G) The numbers of phosphorylated peptides identified by liquid chromatography (LC)-MS/MS analysis of affinity-purified HP1α. (H) Summary of the phosphorylation sites of mouse HP1α deduced from the experiments described above.
    Shrimp Alkaline Phosphatase, supplied by TaKaRa, 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/shrimp alkaline phosphatase/product/TaKaRa
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    shrimp alkaline phosphatase - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

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    Identification of phosphorylation sites of mouse HP1α. (A) The phosphorylation patterns of mouse HP1α analyzed by phos-tag Western blotting (phos-tag WB). Whole-cell lysates (WCLs) of NIH 3T3 cells were resolved on an SDS-PAGE gel containing phos-tag acrylamide (phos-tag) or on a standard SDS-PAGE gel (mock). As an unphosphorylated control, SAP-treated WCL was loaded on the same gel. HP1α was detected using an anti-HP1α antibody. (B) Phosphorylation patterns of mouse HP1α in cells arrested at different cell cycle stages. WCLs were prepared from NIH 3T3 cells grown under the indicated conditions and analyzed by phos-tag WB. The arrowhead indicates an HP1α with basal band shift. The asterisk represents an HP1α species with metaphase-associated phosphorylation. The same membrane was immunoblotted with anti-α-tubulin and anti-TIF1β antibodies for loading controls. (C) Amino acid sequence of mouse HP1α. Light- and dark-shaded boxes indicate the CD and CSD, respectively. The sites of serine (S)-to-alanine (A) substitutions introduced in the following experiments are shown in boldface. (D) Phos-tag and standard WB analysis of wild-type and mutant HP1αs. WCLs were prepared from NIH 3T3 cells transiently expressing FLAG-tagged wild-type HP1α or HP1α with N-terminal (N-term: S11-14A) mutations, hinge (Hinge: S92, 93, 95, 97A) mutations, or both (N-term + hinge). Each FLAG-HP1α with an SAP-treated control was analyzed by phos-tag WB using an anti-FLAG antibody. The phosphorylation state of HP1α represented by each protein band was deduced from the distance of migration level of each band (Rf) and is indicated to the right of the phos-tag WB panel. (E) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the N-terminal region. The phosphorylation patterns of the indicated FLAG-HP1α species were analyzed by phos-tag WB using a higher concentration (80 μM) of phos-tag acrylamide. (F) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the hinge region. (G) The numbers of phosphorylated peptides identified by liquid chromatography (LC)-MS/MS analysis of affinity-purified HP1α. (H) Summary of the phosphorylation sites of mouse HP1α deduced from the experiments described above.

    Journal: Molecular and Cellular Biology

    Article Title: N-Terminal Phosphorylation of HP1? Promotes Its Chromatin Binding ▿

    doi: 10.1128/MCB.01012-10

    Figure Lengend Snippet: Identification of phosphorylation sites of mouse HP1α. (A) The phosphorylation patterns of mouse HP1α analyzed by phos-tag Western blotting (phos-tag WB). Whole-cell lysates (WCLs) of NIH 3T3 cells were resolved on an SDS-PAGE gel containing phos-tag acrylamide (phos-tag) or on a standard SDS-PAGE gel (mock). As an unphosphorylated control, SAP-treated WCL was loaded on the same gel. HP1α was detected using an anti-HP1α antibody. (B) Phosphorylation patterns of mouse HP1α in cells arrested at different cell cycle stages. WCLs were prepared from NIH 3T3 cells grown under the indicated conditions and analyzed by phos-tag WB. The arrowhead indicates an HP1α with basal band shift. The asterisk represents an HP1α species with metaphase-associated phosphorylation. The same membrane was immunoblotted with anti-α-tubulin and anti-TIF1β antibodies for loading controls. (C) Amino acid sequence of mouse HP1α. Light- and dark-shaded boxes indicate the CD and CSD, respectively. The sites of serine (S)-to-alanine (A) substitutions introduced in the following experiments are shown in boldface. (D) Phos-tag and standard WB analysis of wild-type and mutant HP1αs. WCLs were prepared from NIH 3T3 cells transiently expressing FLAG-tagged wild-type HP1α or HP1α with N-terminal (N-term: S11-14A) mutations, hinge (Hinge: S92, 93, 95, 97A) mutations, or both (N-term + hinge). Each FLAG-HP1α with an SAP-treated control was analyzed by phos-tag WB using an anti-FLAG antibody. The phosphorylation state of HP1α represented by each protein band was deduced from the distance of migration level of each band (Rf) and is indicated to the right of the phos-tag WB panel. (E) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the N-terminal region. The phosphorylation patterns of the indicated FLAG-HP1α species were analyzed by phos-tag WB using a higher concentration (80 μM) of phos-tag acrylamide. (F) Phosphorylation state of FLAG-HP1α with each single amino acid substitution in the hinge region. (G) The numbers of phosphorylated peptides identified by liquid chromatography (LC)-MS/MS analysis of affinity-purified HP1α. (H) Summary of the phosphorylation sites of mouse HP1α deduced from the experiments described above.

    Article Snippet: For dephosphorylation, an aliquot of whole-cell lysate (WCL) was incubated with 0.08 U/ml shrimp alkaline phosphatase (SAP) (TaKaRa) for 3 to 6 h at 37°C.

    Techniques: Western Blot, SDS Page, Electrophoretic Mobility Shift Assay, Sequencing, Mutagenesis, Expressing, Migration, Concentration Assay, Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Affinity Purification