pcp biotin  (Jena Bioscience)


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    Name:
    pCp Biotin
    Description:

    Catalog Number:
    NU-1706-BIO
    Price:
    130.3
    Category:
    Nucleotides Nucleosides
    Size:
    20 µl
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    Structured Review

    Jena Bioscience pcp biotin
    Visualization of RBP:RNA complexes with biotin-labeling. (a) (top) Schematic of RBP:RNA visualization experiments, in which three samples are subjected to immunoprecipitation: crosslinked cells with standard (40U) RNase, crosslinked cells with high (333U) RNase, and non-crosslinked cells with either standard or high RNase. RNA was then labeled either through radiolabeling with <t>T4</t> PNK and [γ-32P]-ATP followed by autoradiography, or with T4 RNA Ligase and <t>pCp-Biotin</t> followed by chemiluminescent imaging with streptavidin-conjugated horseradish peroxidase. (bottom) Example RNA imaging with 32 P and biotin-labeling after TIAL1 immunoprecipitation, with standard IP-western shown below. (b-c) Biotin-based RNA labeling for (b) FUS in HepG2 and (c) HNRNPC in K562. (d) Bars indicate the fold-enrichment in biotin-labeled RNA signal between crosslinked versus non-crosslinked samples (with 40U RNase) for the size range from which RNA is isolated (from protein to 75 kDa above). Shown is data from membrane I. Quantification was performed in ImageJ. (e-f) Biotin-based RNA labeling for (e) PTBP1 in HepG2 and (f) NIP7 in HepG2. For each, immunoprecipitated sample was labeled with pCp-Biotin and split in half, with one half transferred to nitrocellulose membrane from supplier I, and the other half transferred to nitrocellulose membrane from supplier G. (right) IP-western experiment from the paired eCLIP experiments). (g) Density plot indicates the number of eCLIP peaks for MATR3 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (h) Bars indicate the fraction of significantly-enriched peaks in ENCODE replicate 2 (performed with membrane I) that are also significantly enriched in (black) ENCODE replicate 1 (membrane I) or (gray) replicates 1 or 2 of a new eCLIP experiment performed with membrane G in the same cell type with the same antibody. (i) Density plot indicates the number of eCLIP peaks for FXR2 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (j) Bars indicate the fold-enrichment for read density at (white) sense or (gray) antisense L1 elements in MATR3 eCLIP in HepG2.

    https://www.bioz.com/result/pcp biotin/product/Jena Bioscience
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcp biotin - by Bioz Stars, 2021-06
    93/100 stars

    Images

    1) Product Images from "Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins"

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins

    Journal: bioRxiv

    doi: 10.1101/807008

    Visualization of RBP:RNA complexes with biotin-labeling. (a) (top) Schematic of RBP:RNA visualization experiments, in which three samples are subjected to immunoprecipitation: crosslinked cells with standard (40U) RNase, crosslinked cells with high (333U) RNase, and non-crosslinked cells with either standard or high RNase. RNA was then labeled either through radiolabeling with T4 PNK and [γ-32P]-ATP followed by autoradiography, or with T4 RNA Ligase and pCp-Biotin followed by chemiluminescent imaging with streptavidin-conjugated horseradish peroxidase. (bottom) Example RNA imaging with 32 P and biotin-labeling after TIAL1 immunoprecipitation, with standard IP-western shown below. (b-c) Biotin-based RNA labeling for (b) FUS in HepG2 and (c) HNRNPC in K562. (d) Bars indicate the fold-enrichment in biotin-labeled RNA signal between crosslinked versus non-crosslinked samples (with 40U RNase) for the size range from which RNA is isolated (from protein to 75 kDa above). Shown is data from membrane I. Quantification was performed in ImageJ. (e-f) Biotin-based RNA labeling for (e) PTBP1 in HepG2 and (f) NIP7 in HepG2. For each, immunoprecipitated sample was labeled with pCp-Biotin and split in half, with one half transferred to nitrocellulose membrane from supplier I, and the other half transferred to nitrocellulose membrane from supplier G. (right) IP-western experiment from the paired eCLIP experiments). (g) Density plot indicates the number of eCLIP peaks for MATR3 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (h) Bars indicate the fraction of significantly-enriched peaks in ENCODE replicate 2 (performed with membrane I) that are also significantly enriched in (black) ENCODE replicate 1 (membrane I) or (gray) replicates 1 or 2 of a new eCLIP experiment performed with membrane G in the same cell type with the same antibody. (i) Density plot indicates the number of eCLIP peaks for FXR2 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (j) Bars indicate the fold-enrichment for read density at (white) sense or (gray) antisense L1 elements in MATR3 eCLIP in HepG2.
    Figure Legend Snippet: Visualization of RBP:RNA complexes with biotin-labeling. (a) (top) Schematic of RBP:RNA visualization experiments, in which three samples are subjected to immunoprecipitation: crosslinked cells with standard (40U) RNase, crosslinked cells with high (333U) RNase, and non-crosslinked cells with either standard or high RNase. RNA was then labeled either through radiolabeling with T4 PNK and [γ-32P]-ATP followed by autoradiography, or with T4 RNA Ligase and pCp-Biotin followed by chemiluminescent imaging with streptavidin-conjugated horseradish peroxidase. (bottom) Example RNA imaging with 32 P and biotin-labeling after TIAL1 immunoprecipitation, with standard IP-western shown below. (b-c) Biotin-based RNA labeling for (b) FUS in HepG2 and (c) HNRNPC in K562. (d) Bars indicate the fold-enrichment in biotin-labeled RNA signal between crosslinked versus non-crosslinked samples (with 40U RNase) for the size range from which RNA is isolated (from protein to 75 kDa above). Shown is data from membrane I. Quantification was performed in ImageJ. (e-f) Biotin-based RNA labeling for (e) PTBP1 in HepG2 and (f) NIP7 in HepG2. For each, immunoprecipitated sample was labeled with pCp-Biotin and split in half, with one half transferred to nitrocellulose membrane from supplier I, and the other half transferred to nitrocellulose membrane from supplier G. (right) IP-western experiment from the paired eCLIP experiments). (g) Density plot indicates the number of eCLIP peaks for MATR3 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (h) Bars indicate the fraction of significantly-enriched peaks in ENCODE replicate 2 (performed with membrane I) that are also significantly enriched in (black) ENCODE replicate 1 (membrane I) or (gray) replicates 1 or 2 of a new eCLIP experiment performed with membrane G in the same cell type with the same antibody. (i) Density plot indicates the number of eCLIP peaks for FXR2 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (j) Bars indicate the fold-enrichment for read density at (white) sense or (gray) antisense L1 elements in MATR3 eCLIP in HepG2.

    Techniques Used: Labeling, Immunoprecipitation, Radioactivity, Autoradiography, Imaging, Western Blot, Isolation

    Related Articles

    In Vitro:

    Article Title: Metabolome and transcriptome-wide effects of the carbon storage regulator A in enteropathogenic Escherichia coli
    Article Snippet: RNA was synthesized in vitro using the TranscriptionAid T7 High Yield Transcription Kit (Fermentas) and purified by phenol purification as described elsewhere. .. In vitro transcribed sRNA was biotin-labeled using 6 µl of 10x RNA-Ligase buffer (New England Biolabs (NEB)), 6 µl 10 mM ATP (NEB), 2 µg wza RNA, 6 µl DMSO (NEB), 6 µl 10 µM pCp-biotin (Jena Bioscience) and 18 µl 50% PEG-8000 (NEB) in a total volume of 60 µl at 17 °C overnight. .. RNA was incubated for 10 min at 70 °C before addition to the reaction mix.

    Incubation:

    Article Title: Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone
    Article Snippet: The purified RNAs were heated to 95°C for 5 min and chilled on ice. .. A 25 µL biotinylation reaction contained 1× T4 RNA ligase buffer (NEB), 1 µM RNA, 2 µM biotin-pCp (Jena Bioscience), 10% DMSO, 16% (w/v) PEG-8000, 1 mM ATP, 32.5 U RNasin (Promega), and 50 U T4-RNA ligase I (NEB) and was incubated for 16 h at 6°C. .. For 3′-biotinylated nLuc RNAs, 7 mM anti-reverse cap analog (NEB) was used during transcription.

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins
    Article Snippet: .. At this point, a modified RNA linker ligation was performed with standard eCLIP ligation conditions (buffer and High Concentration T4 RNA Ligase) but with 500 pmol pCp-Biotin (Jena Bioscience) in place of the RNA adapter, and samples were incubated at 16°C. .. For some experiments, immunoprecipitations were performed on 4 million cells; for these experiments, half reactions were used for the pCp-biotin ligation step.

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins
    Article Snippet: At this point, a modified RNA linker ligation was performed with standard eCLIP ligation conditions (buffer and High ?A3B2 show $132#? .. > Concentration T4 RNA Ligase) but with 500 pmol pCp-Biotin (Jena Bioscience) in place of the RNA adapter, and samples were incubated at 16 °C. .. For some experiments, immunoprecipitations were performed on 4 million cells; for these experiments, half reactions were used for the pCp-biotin ligation step.

    Modification:

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins
    Article Snippet: .. At this point, a modified RNA linker ligation was performed with standard eCLIP ligation conditions (buffer and High Concentration T4 RNA Ligase) but with 500 pmol pCp-Biotin (Jena Bioscience) in place of the RNA adapter, and samples were incubated at 16°C. .. For some experiments, immunoprecipitations were performed on 4 million cells; for these experiments, half reactions were used for the pCp-biotin ligation step.

    Ligation:

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins
    Article Snippet: .. At this point, a modified RNA linker ligation was performed with standard eCLIP ligation conditions (buffer and High Concentration T4 RNA Ligase) but with 500 pmol pCp-Biotin (Jena Bioscience) in place of the RNA adapter, and samples were incubated at 16°C. .. For some experiments, immunoprecipitations were performed on 4 million cells; for these experiments, half reactions were used for the pCp-biotin ligation step.

    Article Title: A bacterial secreted translocator hijacks riboregulators to control type III secretion in response to host cell contact
    Article Snippet: Finally, the RNA run-off transcript was purified by phenol-chloroform extraction [ ]. .. RNA electrophoretic mobility shift assay (RNA-EMSA) For the RNA-EMSAs, some RNAs were 3’-end labeled with pCp Biotin (Jena, Bioscience, Germany) by ligation with T4 RNA Ligase I (lcrF transcripts, ; csrA (a) transcript, ; csrA transcript (e) and (f), ). .. The reaction was performed with 1 μg linearized RNA in 1 x T4 ligase buffer, 1 mM ATP, 10% (w/v) DMSO, 1 μM pCp-Biotin (Jena, Bioscience, Germany), 15% (w/v) PEG8000 and 1 μl T4 ssRNA ligase (20 u, NEB, USA) at 18°C for 2 h or overnight.

    Concentration Assay:

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins
    Article Snippet: .. At this point, a modified RNA linker ligation was performed with standard eCLIP ligation conditions (buffer and High Concentration T4 RNA Ligase) but with 500 pmol pCp-Biotin (Jena Bioscience) in place of the RNA adapter, and samples were incubated at 16°C. .. For some experiments, immunoprecipitations were performed on 4 million cells; for these experiments, half reactions were used for the pCp-biotin ligation step.

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins
    Article Snippet: At this point, a modified RNA linker ligation was performed with standard eCLIP ligation conditions (buffer and High ?A3B2 show $132#? .. > Concentration T4 RNA Ligase) but with 500 pmol pCp-Biotin (Jena Bioscience) in place of the RNA adapter, and samples were incubated at 16 °C. .. For some experiments, immunoprecipitations were performed on 4 million cells; for these experiments, half reactions were used for the pCp-biotin ligation step.

    Electrophoretic Mobility Shift Assay:

    Article Title: A bacterial secreted translocator hijacks riboregulators to control type III secretion in response to host cell contact
    Article Snippet: Finally, the RNA run-off transcript was purified by phenol-chloroform extraction [ ]. .. RNA electrophoretic mobility shift assay (RNA-EMSA) For the RNA-EMSAs, some RNAs were 3’-end labeled with pCp Biotin (Jena, Bioscience, Germany) by ligation with T4 RNA Ligase I (lcrF transcripts, ; csrA (a) transcript, ; csrA transcript (e) and (f), ). .. The reaction was performed with 1 μg linearized RNA in 1 x T4 ligase buffer, 1 mM ATP, 10% (w/v) DMSO, 1 μM pCp-Biotin (Jena, Bioscience, Germany), 15% (w/v) PEG8000 and 1 μl T4 ssRNA ligase (20 u, NEB, USA) at 18°C for 2 h or overnight.

    Labeling:

    Article Title: A bacterial secreted translocator hijacks riboregulators to control type III secretion in response to host cell contact
    Article Snippet: Finally, the RNA run-off transcript was purified by phenol-chloroform extraction [ ]. .. RNA electrophoretic mobility shift assay (RNA-EMSA) For the RNA-EMSAs, some RNAs were 3’-end labeled with pCp Biotin (Jena, Bioscience, Germany) by ligation with T4 RNA Ligase I (lcrF transcripts, ; csrA (a) transcript, ; csrA transcript (e) and (f), ). .. The reaction was performed with 1 μg linearized RNA in 1 x T4 ligase buffer, 1 mM ATP, 10% (w/v) DMSO, 1 μM pCp-Biotin (Jena, Bioscience, Germany), 15% (w/v) PEG8000 and 1 μl T4 ssRNA ligase (20 u, NEB, USA) at 18°C for 2 h or overnight.

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    Jena Bioscience pcp biotin
    Visualization of RBP:RNA complexes with biotin-labeling. (a) (top) Schematic of RBP:RNA visualization experiments, in which three samples are subjected to immunoprecipitation: crosslinked cells with standard (40U) RNase, crosslinked cells with high (333U) RNase, and non-crosslinked cells with either standard or high RNase. RNA was then labeled either through radiolabeling with <t>T4</t> PNK and [γ-32P]-ATP followed by autoradiography, or with T4 RNA Ligase and <t>pCp-Biotin</t> followed by chemiluminescent imaging with streptavidin-conjugated horseradish peroxidase. (bottom) Example RNA imaging with 32 P and biotin-labeling after TIAL1 immunoprecipitation, with standard IP-western shown below. (b-c) Biotin-based RNA labeling for (b) FUS in HepG2 and (c) HNRNPC in K562. (d) Bars indicate the fold-enrichment in biotin-labeled RNA signal between crosslinked versus non-crosslinked samples (with 40U RNase) for the size range from which RNA is isolated (from protein to 75 kDa above). Shown is data from membrane I. Quantification was performed in ImageJ. (e-f) Biotin-based RNA labeling for (e) PTBP1 in HepG2 and (f) NIP7 in HepG2. For each, immunoprecipitated sample was labeled with pCp-Biotin and split in half, with one half transferred to nitrocellulose membrane from supplier I, and the other half transferred to nitrocellulose membrane from supplier G. (right) IP-western experiment from the paired eCLIP experiments). (g) Density plot indicates the number of eCLIP peaks for MATR3 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (h) Bars indicate the fraction of significantly-enriched peaks in ENCODE replicate 2 (performed with membrane I) that are also significantly enriched in (black) ENCODE replicate 1 (membrane I) or (gray) replicates 1 or 2 of a new eCLIP experiment performed with membrane G in the same cell type with the same antibody. (i) Density plot indicates the number of eCLIP peaks for FXR2 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (j) Bars indicate the fold-enrichment for read density at (white) sense or (gray) antisense L1 elements in MATR3 eCLIP in HepG2.
    Pcp Biotin, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcp biotin/product/Jena Bioscience
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcp biotin - by Bioz Stars, 2021-06
    93/100 stars
      Buy from Supplier

    Image Search Results


    Visualization of RBP:RNA complexes with biotin-labeling. (a) (top) Schematic of RBP:RNA visualization experiments, in which three samples are subjected to immunoprecipitation: crosslinked cells with standard (40U) RNase, crosslinked cells with high (333U) RNase, and non-crosslinked cells with either standard or high RNase. RNA was then labeled either through radiolabeling with T4 PNK and [γ-32P]-ATP followed by autoradiography, or with T4 RNA Ligase and pCp-Biotin followed by chemiluminescent imaging with streptavidin-conjugated horseradish peroxidase. (bottom) Example RNA imaging with 32 P and biotin-labeling after TIAL1 immunoprecipitation, with standard IP-western shown below. (b-c) Biotin-based RNA labeling for (b) FUS in HepG2 and (c) HNRNPC in K562. (d) Bars indicate the fold-enrichment in biotin-labeled RNA signal between crosslinked versus non-crosslinked samples (with 40U RNase) for the size range from which RNA is isolated (from protein to 75 kDa above). Shown is data from membrane I. Quantification was performed in ImageJ. (e-f) Biotin-based RNA labeling for (e) PTBP1 in HepG2 and (f) NIP7 in HepG2. For each, immunoprecipitated sample was labeled with pCp-Biotin and split in half, with one half transferred to nitrocellulose membrane from supplier I, and the other half transferred to nitrocellulose membrane from supplier G. (right) IP-western experiment from the paired eCLIP experiments). (g) Density plot indicates the number of eCLIP peaks for MATR3 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (h) Bars indicate the fraction of significantly-enriched peaks in ENCODE replicate 2 (performed with membrane I) that are also significantly enriched in (black) ENCODE replicate 1 (membrane I) or (gray) replicates 1 or 2 of a new eCLIP experiment performed with membrane G in the same cell type with the same antibody. (i) Density plot indicates the number of eCLIP peaks for FXR2 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (j) Bars indicate the fold-enrichment for read density at (white) sense or (gray) antisense L1 elements in MATR3 eCLIP in HepG2.

    Journal: bioRxiv

    Article Title: Principles of RNA processing from analysis of enhanced CLIP maps for 150 RNA binding proteins

    doi: 10.1101/807008

    Figure Lengend Snippet: Visualization of RBP:RNA complexes with biotin-labeling. (a) (top) Schematic of RBP:RNA visualization experiments, in which three samples are subjected to immunoprecipitation: crosslinked cells with standard (40U) RNase, crosslinked cells with high (333U) RNase, and non-crosslinked cells with either standard or high RNase. RNA was then labeled either through radiolabeling with T4 PNK and [γ-32P]-ATP followed by autoradiography, or with T4 RNA Ligase and pCp-Biotin followed by chemiluminescent imaging with streptavidin-conjugated horseradish peroxidase. (bottom) Example RNA imaging with 32 P and biotin-labeling after TIAL1 immunoprecipitation, with standard IP-western shown below. (b-c) Biotin-based RNA labeling for (b) FUS in HepG2 and (c) HNRNPC in K562. (d) Bars indicate the fold-enrichment in biotin-labeled RNA signal between crosslinked versus non-crosslinked samples (with 40U RNase) for the size range from which RNA is isolated (from protein to 75 kDa above). Shown is data from membrane I. Quantification was performed in ImageJ. (e-f) Biotin-based RNA labeling for (e) PTBP1 in HepG2 and (f) NIP7 in HepG2. For each, immunoprecipitated sample was labeled with pCp-Biotin and split in half, with one half transferred to nitrocellulose membrane from supplier I, and the other half transferred to nitrocellulose membrane from supplier G. (right) IP-western experiment from the paired eCLIP experiments). (g) Density plot indicates the number of eCLIP peaks for MATR3 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (h) Bars indicate the fraction of significantly-enriched peaks in ENCODE replicate 2 (performed with membrane I) that are also significantly enriched in (black) ENCODE replicate 1 (membrane I) or (gray) replicates 1 or 2 of a new eCLIP experiment performed with membrane G in the same cell type with the same antibody. (i) Density plot indicates the number of eCLIP peaks for FXR2 in HepG2 identified as significant in ENCODE replicate 2 that have the indicated fold-enrichment in (x-axis) ENCODE replicate 1 (performed with membrane I) versus (y-axis) a new eCLIP replicate performed with membrane G. Color indicates the number of points within each hexagon. (j) Bars indicate the fold-enrichment for read density at (white) sense or (gray) antisense L1 elements in MATR3 eCLIP in HepG2.

    Article Snippet: At this point, a modified RNA linker ligation was performed with standard eCLIP ligation conditions (buffer and High Concentration T4 RNA Ligase) but with 500 pmol pCp-Biotin (Jena Bioscience) in place of the RNA adapter, and samples were incubated at 16°C.

    Techniques: Labeling, Immunoprecipitation, Radioactivity, Autoradiography, Imaging, Western Blot, Isolation