anti brutp  (Roche)


Bioz Verified Symbol Roche is a verified supplier
Bioz Manufacturer Symbol Roche manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85

    Structured Review

    Roche anti brutp
    (A) NF-Y is associated with active transcription sites in living cells. After in vivo incorporation of <t>BrUTP</t> (run-on), cells were fixed and endogenous NF-YA (ii) and nascent RNA transcripts (i) were detected by indirect immunofluorescence combined with Confocal Scanning Laser Microscopy by using anti-NF-YA and anti-BrU antibodies. In the overlay (iii), yellow indicates colocalizations between NF-YA (green) and transcription sites (red). In panels vi and x cells were immunostained with anti-NF-YA, in panels v and ix with <t>anti-RPII</t> CTD repeat YSPTSPS (phospho S2) and anti-total RPII respectively. The majority of NF-YA (red) colocalizes with the activated form of RPII (green)(vii). (B) Cells were immunostained with anti-NF-YA (vii-xii), -acetylated H3K9 (i), -acetylated H4 (ii), -tri-methylated H3K9 (iii), -di-methylated H3K27 (iv), -tri-methylated H4K20 (v) and -NF-YB (vi) antibodies. The majority of NF-YA colocalizes with acetylated (xiii, xiv), but poor colocalization occurs with methylated histones (xv, xvi,xvii). Panel xviii shows the overlay of two subunits of NF-Y, NF-YA (xii) and NF-YB (vi). Panels from xix to xxiv represent a typical optical field of the merge. In figure 1A and 1B confocal analysis of single optical section is shown. The images have been collected with a 60x objective.
    Anti Brutp, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 292 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti brutp/product/Roche
    Average 85 stars, based on 292 article reviews
    Price from $9.99 to $1999.99
    anti brutp - by Bioz Stars, 2020-09
    85/100 stars

    Images

    1) Product Images from "NF-Y Dependent Epigenetic Modifications Discriminate between Proliferating and Postmitotic Tissue"

    Article Title: NF-Y Dependent Epigenetic Modifications Discriminate between Proliferating and Postmitotic Tissue

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0002047

    (A) NF-Y is associated with active transcription sites in living cells. After in vivo incorporation of BrUTP (run-on), cells were fixed and endogenous NF-YA (ii) and nascent RNA transcripts (i) were detected by indirect immunofluorescence combined with Confocal Scanning Laser Microscopy by using anti-NF-YA and anti-BrU antibodies. In the overlay (iii), yellow indicates colocalizations between NF-YA (green) and transcription sites (red). In panels vi and x cells were immunostained with anti-NF-YA, in panels v and ix with anti-RPII CTD repeat YSPTSPS (phospho S2) and anti-total RPII respectively. The majority of NF-YA (red) colocalizes with the activated form of RPII (green)(vii). (B) Cells were immunostained with anti-NF-YA (vii-xii), -acetylated H3K9 (i), -acetylated H4 (ii), -tri-methylated H3K9 (iii), -di-methylated H3K27 (iv), -tri-methylated H4K20 (v) and -NF-YB (vi) antibodies. The majority of NF-YA colocalizes with acetylated (xiii, xiv), but poor colocalization occurs with methylated histones (xv, xvi,xvii). Panel xviii shows the overlay of two subunits of NF-Y, NF-YA (xii) and NF-YB (vi). Panels from xix to xxiv represent a typical optical field of the merge. In figure 1A and 1B confocal analysis of single optical section is shown. The images have been collected with a 60x objective.
    Figure Legend Snippet: (A) NF-Y is associated with active transcription sites in living cells. After in vivo incorporation of BrUTP (run-on), cells were fixed and endogenous NF-YA (ii) and nascent RNA transcripts (i) were detected by indirect immunofluorescence combined with Confocal Scanning Laser Microscopy by using anti-NF-YA and anti-BrU antibodies. In the overlay (iii), yellow indicates colocalizations between NF-YA (green) and transcription sites (red). In panels vi and x cells were immunostained with anti-NF-YA, in panels v and ix with anti-RPII CTD repeat YSPTSPS (phospho S2) and anti-total RPII respectively. The majority of NF-YA (red) colocalizes with the activated form of RPII (green)(vii). (B) Cells were immunostained with anti-NF-YA (vii-xii), -acetylated H3K9 (i), -acetylated H4 (ii), -tri-methylated H3K9 (iii), -di-methylated H3K27 (iv), -tri-methylated H4K20 (v) and -NF-YB (vi) antibodies. The majority of NF-YA colocalizes with acetylated (xiii, xiv), but poor colocalization occurs with methylated histones (xv, xvi,xvii). Panel xviii shows the overlay of two subunits of NF-Y, NF-YA (xii) and NF-YB (vi). Panels from xix to xxiv represent a typical optical field of the merge. In figure 1A and 1B confocal analysis of single optical section is shown. The images have been collected with a 60x objective.

    Techniques Used: In Vivo, Immunofluorescence, Microscopy, Methylation

    2) Product Images from "Membrane Association and Dimerization of a Cysteine-Rich, 16-Kilodalton Polypeptide Released from the C-Terminal Region of the Coronavirus Infectious Bronchitis Virus 1a Polyprotein"

    Article Title: Membrane Association and Dimerization of a Cysteine-Rich, 16-Kilodalton Polypeptide Released from the C-Terminal Region of the Coronavirus Infectious Bronchitis Virus 1a Polyprotein

    Journal: Journal of Virology

    doi: 10.1128/JVI.76.12.6257-6267.2002

    (a) Subcellular localization of the 16-kDa protein. (A) Cos-7 cells overexpressing pEGFP-16k at 16 h posttransfection showing perinuclear staining. (B) Cos-7 cells overexpressing the EGFP control vector (pEGFP) showing diffuse staining. (C) Immunofluorescent staining of IBV-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection showing distinct punctate perinuclear staining. (D) Immunofluorescent staining of mock-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection. (E) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining profile of anti-16-kDa protein serum. (F) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining pattern of anti-BrUTP serum. (G) Superimposition of images E and F. All images were taken from a Zeiss Axioplan confocal microscope. (b) Membrane association of the 16-kDa protein. Cos-7 cells expressing the 16-kDa protein were labeled with [ 35 S]methionine-cysteine for 4 h and harvested. Cells were lysed with a Dounce homogenizer and fractionated into membrane (M) and cytosol (C) fractions at pH 7 (lanes 1 and 2) by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography. Membrane (M) pellets were resuspended in hypotonic buffer; treated with 1% Triton X-100 (TX-100), 100 mM Na 2 CO 3 , or 1 M KCl; and further fractionated into supernatant (S) and pellet (P) fractions by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography.
    Figure Legend Snippet: (a) Subcellular localization of the 16-kDa protein. (A) Cos-7 cells overexpressing pEGFP-16k at 16 h posttransfection showing perinuclear staining. (B) Cos-7 cells overexpressing the EGFP control vector (pEGFP) showing diffuse staining. (C) Immunofluorescent staining of IBV-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection showing distinct punctate perinuclear staining. (D) Immunofluorescent staining of mock-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection. (E) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining profile of anti-16-kDa protein serum. (F) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining pattern of anti-BrUTP serum. (G) Superimposition of images E and F. All images were taken from a Zeiss Axioplan confocal microscope. (b) Membrane association of the 16-kDa protein. Cos-7 cells expressing the 16-kDa protein were labeled with [ 35 S]methionine-cysteine for 4 h and harvested. Cells were lysed with a Dounce homogenizer and fractionated into membrane (M) and cytosol (C) fractions at pH 7 (lanes 1 and 2) by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography. Membrane (M) pellets were resuspended in hypotonic buffer; treated with 1% Triton X-100 (TX-100), 100 mM Na 2 CO 3 , or 1 M KCl; and further fractionated into supernatant (S) and pellet (P) fractions by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography.

    Techniques Used: Staining, Plasmid Preparation, Infection, Labeling, Transfection, Microscopy, Expressing, Immunoprecipitation

    3) Product Images from "Membrane Association and Dimerization of a Cysteine-Rich, 16-Kilodalton Polypeptide Released from the C-Terminal Region of the Coronavirus Infectious Bronchitis Virus 1a Polyprotein"

    Article Title: Membrane Association and Dimerization of a Cysteine-Rich, 16-Kilodalton Polypeptide Released from the C-Terminal Region of the Coronavirus Infectious Bronchitis Virus 1a Polyprotein

    Journal: Journal of Virology

    doi: 10.1128/JVI.76.12.6257-6267.2002

    (a) Subcellular localization of the 16-kDa protein. (A) Cos-7 cells overexpressing pEGFP-16k at 16 h posttransfection showing perinuclear staining. (B) Cos-7 cells overexpressing the EGFP control vector (pEGFP) showing diffuse staining. (C) Immunofluorescent staining of IBV-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection showing distinct punctate perinuclear staining. (D) Immunofluorescent staining of mock-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection. (E) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining profile of anti-16-kDa protein serum. (F) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining pattern of anti-BrUTP serum. (G) Superimposition of images E and F. All images were taken from a Zeiss Axioplan confocal microscope. (b) Membrane association of the 16-kDa protein. Cos-7 cells expressing the 16-kDa protein were labeled with [ 35 S]methionine-cysteine for 4 h and harvested. Cells were lysed with a Dounce homogenizer and fractionated into membrane (M) and cytosol (C) fractions at pH 7 (lanes 1 and 2) by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography. Membrane (M) pellets were resuspended in hypotonic buffer; treated with 1% Triton X-100 (TX-100), 100 mM Na 2 CO 3 , or 1 M KCl; and further fractionated into supernatant (S) and pellet (P) fractions by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography.
    Figure Legend Snippet: (a) Subcellular localization of the 16-kDa protein. (A) Cos-7 cells overexpressing pEGFP-16k at 16 h posttransfection showing perinuclear staining. (B) Cos-7 cells overexpressing the EGFP control vector (pEGFP) showing diffuse staining. (C) Immunofluorescent staining of IBV-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection showing distinct punctate perinuclear staining. (D) Immunofluorescent staining of mock-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection. (E) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining profile of anti-16-kDa protein serum. (F) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining pattern of anti-BrUTP serum. (G) Superimposition of images E and F. All images were taken from a Zeiss Axioplan confocal microscope. (b) Membrane association of the 16-kDa protein. Cos-7 cells expressing the 16-kDa protein were labeled with [ 35 S]methionine-cysteine for 4 h and harvested. Cells were lysed with a Dounce homogenizer and fractionated into membrane (M) and cytosol (C) fractions at pH 7 (lanes 1 and 2) by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography. Membrane (M) pellets were resuspended in hypotonic buffer; treated with 1% Triton X-100 (TX-100), 100 mM Na 2 CO 3 , or 1 M KCl; and further fractionated into supernatant (S) and pellet (P) fractions by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography.

    Techniques Used: Staining, Plasmid Preparation, Infection, Labeling, Transfection, Microscopy, Expressing, Immunoprecipitation

    (a) Diagram of ORFs 1a and 1b illustrating the two overlapping papain-like proteinase domains (PLPDs), the 3C-like proteinase (3CLpro), the RNA-dependent RNA polymerase, the metal-binding domain, and the helicase. The Q 3783 -S 3784 and Q 3928 -S 3929 ) of the 16-kDa protein. The molecular mass of the 16-kDa protein was estimated on the basis of its migration on SDS-PAGE. The Q 3783 -S 3784 and Q 3928 -S 3929 residues are encoded by nucleotides 11,875 to 11,880 and 12,310 to 12,315, respectively. The 12 cysteine residues are in bold. (b) Detection of the 16-kDa protein in IBV-infected Vero cells. IBV-infected (IBV) and mock-infected (Mock) Vero cells were labeled with [ 35 S]methionine-cysteine and harvested at 12 h postinfection. The cell debris and nuclei fractions were removed by low-speed centrifugation (1,500 × g ) for 10 min, and the cytosol and membrane fractions were separated by high-speed centrifugation (150,000 × g ). The cytosol and membrane fractions were immunoprecipitated with anti-16-kDa protein serum and anti-M serum. Polypeptides were separated on an SDS-15% polyacrylamide gel and detected by fluorography. The values on the left are molecular sizes in kilodaltons.
    Figure Legend Snippet: (a) Diagram of ORFs 1a and 1b illustrating the two overlapping papain-like proteinase domains (PLPDs), the 3C-like proteinase (3CLpro), the RNA-dependent RNA polymerase, the metal-binding domain, and the helicase. The Q 3783 -S 3784 and Q 3928 -S 3929 ) of the 16-kDa protein. The molecular mass of the 16-kDa protein was estimated on the basis of its migration on SDS-PAGE. The Q 3783 -S 3784 and Q 3928 -S 3929 residues are encoded by nucleotides 11,875 to 11,880 and 12,310 to 12,315, respectively. The 12 cysteine residues are in bold. (b) Detection of the 16-kDa protein in IBV-infected Vero cells. IBV-infected (IBV) and mock-infected (Mock) Vero cells were labeled with [ 35 S]methionine-cysteine and harvested at 12 h postinfection. The cell debris and nuclei fractions were removed by low-speed centrifugation (1,500 × g ) for 10 min, and the cytosol and membrane fractions were separated by high-speed centrifugation (150,000 × g ). The cytosol and membrane fractions were immunoprecipitated with anti-16-kDa protein serum and anti-M serum. Polypeptides were separated on an SDS-15% polyacrylamide gel and detected by fluorography. The values on the left are molecular sizes in kilodaltons.

    Techniques Used: Binding Assay, Migration, SDS Page, Infection, Labeling, Centrifugation, Immunoprecipitation

    4) Product Images from "Membrane Association and Dimerization of a Cysteine-Rich, 16-Kilodalton Polypeptide Released from the C-Terminal Region of the Coronavirus Infectious Bronchitis Virus 1a Polyprotein"

    Article Title: Membrane Association and Dimerization of a Cysteine-Rich, 16-Kilodalton Polypeptide Released from the C-Terminal Region of the Coronavirus Infectious Bronchitis Virus 1a Polyprotein

    Journal: Journal of Virology

    doi: 10.1128/JVI.76.12.6257-6267.2002

    (a) Effect of EGF on expression of the 16-kDa protein. Cells were transfected by electroporation with RNAs transcribed from plasmids (as indicated above the lanes) and stimulated with 50 ng of EGF per ml for 30 min at 48 h postinfection. Cells were harvested, and gel electrophoresis of total proteins was performed on an SDS-15% polyacrylamide gel. The proteins were transferred to nitrocellulose membranes; blotted with anti-16-kDa protein, anti-GFP, and anti-β-tubulin sera; and detected with an ECL + Plus Western blotting detection kit. (b) Cotransfection of the 16-kDa protein and human EGFR. Cells were cotransfected by electroporation with RNAs transcribed from plasmids pSinRep16k and pSinRepEGFR and stimulated with 50 ng of EGF per ml for 30 min at 48 h posttransfection (lanes 1 and 2). Cells transfected with RNA transcribed from plasmid pSinRepGFP were included as a control. Cells were harvested, and gel electrophoresis of the total proteins was performed on an SDS-15% polyacrylamide gel. The proteins were transferred to nitrocellulose membranes; blotted with anti-phosphorylated EGFR, anti-EGFR, anti-16-kDa protein, and anti-β-tubulin sera; and detected with an ECL + Plus Western blotting detection kit. (c) Effect of EGF stimulation on dimerization of the 16-kDa protein. Cells were transfected by electroporation with RNA transcribed from plasmid pSinRep16k and stimulated with 50 ng of EGF per ml at 48 h posttransfection. Lysates were prepared in TGP buffer, and gel electrophoresis of total proteins was performed on an SDS-15% polyacrylamide gel under either nonreducing (NR) or reducing (R) conditions. The proteins were transferred to nitrocellulose membranes, blotted with anti-16-kDa protein serum and anti-β-tubulin serum, and detected with an ECL + Plus Western blotting detection kit.
    Figure Legend Snippet: (a) Effect of EGF on expression of the 16-kDa protein. Cells were transfected by electroporation with RNAs transcribed from plasmids (as indicated above the lanes) and stimulated with 50 ng of EGF per ml for 30 min at 48 h postinfection. Cells were harvested, and gel electrophoresis of total proteins was performed on an SDS-15% polyacrylamide gel. The proteins were transferred to nitrocellulose membranes; blotted with anti-16-kDa protein, anti-GFP, and anti-β-tubulin sera; and detected with an ECL + Plus Western blotting detection kit. (b) Cotransfection of the 16-kDa protein and human EGFR. Cells were cotransfected by electroporation with RNAs transcribed from plasmids pSinRep16k and pSinRepEGFR and stimulated with 50 ng of EGF per ml for 30 min at 48 h posttransfection (lanes 1 and 2). Cells transfected with RNA transcribed from plasmid pSinRepGFP were included as a control. Cells were harvested, and gel electrophoresis of the total proteins was performed on an SDS-15% polyacrylamide gel. The proteins were transferred to nitrocellulose membranes; blotted with anti-phosphorylated EGFR, anti-EGFR, anti-16-kDa protein, and anti-β-tubulin sera; and detected with an ECL + Plus Western blotting detection kit. (c) Effect of EGF stimulation on dimerization of the 16-kDa protein. Cells were transfected by electroporation with RNA transcribed from plasmid pSinRep16k and stimulated with 50 ng of EGF per ml at 48 h posttransfection. Lysates were prepared in TGP buffer, and gel electrophoresis of total proteins was performed on an SDS-15% polyacrylamide gel under either nonreducing (NR) or reducing (R) conditions. The proteins were transferred to nitrocellulose membranes, blotted with anti-16-kDa protein serum and anti-β-tubulin serum, and detected with an ECL + Plus Western blotting detection kit.

    Techniques Used: Expressing, Transfection, Electroporation, Nucleic Acid Electrophoresis, Western Blot, Cotransfection, Plasmid Preparation

    (a) Subcellular localization of the 16-kDa protein. (A) Cos-7 cells overexpressing pEGFP-16k at 16 h posttransfection showing perinuclear staining. (B) Cos-7 cells overexpressing the EGFP control vector (pEGFP) showing diffuse staining. (C) Immunofluorescent staining of IBV-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection showing distinct punctate perinuclear staining. (D) Immunofluorescent staining of mock-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection. (E) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining profile of anti-16-kDa protein serum. (F) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining pattern of anti-BrUTP serum. (G) Superimposition of images E and F. All images were taken from a Zeiss Axioplan confocal microscope. (b) Membrane association of the 16-kDa protein. Cos-7 cells expressing the 16-kDa protein were labeled with [ 35 S]methionine-cysteine for 4 h and harvested. Cells were lysed with a Dounce homogenizer and fractionated into membrane (M) and cytosol (C) fractions at pH 7 (lanes 1 and 2) by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography. Membrane (M) pellets were resuspended in hypotonic buffer; treated with 1% Triton X-100 (TX-100), 100 mM Na 2 CO 3 , or 1 M KCl; and further fractionated into supernatant (S) and pellet (P) fractions by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography.
    Figure Legend Snippet: (a) Subcellular localization of the 16-kDa protein. (A) Cos-7 cells overexpressing pEGFP-16k at 16 h posttransfection showing perinuclear staining. (B) Cos-7 cells overexpressing the EGFP control vector (pEGFP) showing diffuse staining. (C) Immunofluorescent staining of IBV-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection showing distinct punctate perinuclear staining. (D) Immunofluorescent staining of mock-infected Vero cells with anti-16-kDa protein serum (1:20) at 13 h postinfection. (E) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining profile of anti-16-kDa protein serum. (F) Double labeling of IBV-infected Vero cells that were transfected with BrUTP at 10 h postinfection showing the staining pattern of anti-BrUTP serum. (G) Superimposition of images E and F. All images were taken from a Zeiss Axioplan confocal microscope. (b) Membrane association of the 16-kDa protein. Cos-7 cells expressing the 16-kDa protein were labeled with [ 35 S]methionine-cysteine for 4 h and harvested. Cells were lysed with a Dounce homogenizer and fractionated into membrane (M) and cytosol (C) fractions at pH 7 (lanes 1 and 2) by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography. Membrane (M) pellets were resuspended in hypotonic buffer; treated with 1% Triton X-100 (TX-100), 100 mM Na 2 CO 3 , or 1 M KCl; and further fractionated into supernatant (S) and pellet (P) fractions by ultracentrifugation. Polypeptides were immunoprecipitated with anti-16-kDa protein serum, separated on an SDS-15% polyacrylamide gel, and detected by fluorography.

    Techniques Used: Staining, Plasmid Preparation, Infection, Labeling, Transfection, Microscopy, Expressing, Immunoprecipitation

    (a) Diagram of ORFs 1a and 1b illustrating the two overlapping papain-like proteinase domains (PLPDs), the 3C-like proteinase (3CLpro), the RNA-dependent RNA polymerase, the metal-binding domain, and the helicase. The Q 3783 -S 3784 and Q 3928 -S 3929 ) of the 16-kDa protein. The molecular mass of the 16-kDa protein was estimated on the basis of its migration on SDS-PAGE. The Q 3783 -S 3784 and Q 3928 -S 3929 residues are encoded by nucleotides 11,875 to 11,880 and 12,310 to 12,315, respectively. The 12 cysteine residues are in bold. (b) Detection of the 16-kDa protein in IBV-infected Vero cells. IBV-infected (IBV) and mock-infected (Mock) Vero cells were labeled with [ 35 S]methionine-cysteine and harvested at 12 h postinfection. The cell debris and nuclei fractions were removed by low-speed centrifugation (1,500 × g ) for 10 min, and the cytosol and membrane fractions were separated by high-speed centrifugation (150,000 × g ). The cytosol and membrane fractions were immunoprecipitated with anti-16-kDa protein serum and anti-M serum. Polypeptides were separated on an SDS-15% polyacrylamide gel and detected by fluorography. The values on the left are molecular sizes in kilodaltons.
    Figure Legend Snippet: (a) Diagram of ORFs 1a and 1b illustrating the two overlapping papain-like proteinase domains (PLPDs), the 3C-like proteinase (3CLpro), the RNA-dependent RNA polymerase, the metal-binding domain, and the helicase. The Q 3783 -S 3784 and Q 3928 -S 3929 ) of the 16-kDa protein. The molecular mass of the 16-kDa protein was estimated on the basis of its migration on SDS-PAGE. The Q 3783 -S 3784 and Q 3928 -S 3929 residues are encoded by nucleotides 11,875 to 11,880 and 12,310 to 12,315, respectively. The 12 cysteine residues are in bold. (b) Detection of the 16-kDa protein in IBV-infected Vero cells. IBV-infected (IBV) and mock-infected (Mock) Vero cells were labeled with [ 35 S]methionine-cysteine and harvested at 12 h postinfection. The cell debris and nuclei fractions were removed by low-speed centrifugation (1,500 × g ) for 10 min, and the cytosol and membrane fractions were separated by high-speed centrifugation (150,000 × g ). The cytosol and membrane fractions were immunoprecipitated with anti-16-kDa protein serum and anti-M serum. Polypeptides were separated on an SDS-15% polyacrylamide gel and detected by fluorography. The values on the left are molecular sizes in kilodaltons.

    Techniques Used: Binding Assay, Migration, SDS Page, Infection, Labeling, Centrifugation, Immunoprecipitation

    5) Product Images from "NF-Y Dependent Epigenetic Modifications Discriminate between Proliferating and Postmitotic Tissue"

    Article Title: NF-Y Dependent Epigenetic Modifications Discriminate between Proliferating and Postmitotic Tissue

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0002047

    Lack of NF-Y DNA binding activity induces hypermethylation of histones and target gene inactivation. (A) RT-PCR amplification of the indicated genes from uninfected MEFs (lane 1), MEFs 24hours after infection with Ad-GFP (lane 2), MEFs 24hours and 48hours after infection with Ad-DN-NFYA (lanes 3 and 4). (B) Western blot analysis performed on total lysates from: Mef after 24hours post infection with Ad-GFP (lane1), Mef after 24 hours post infection with Ad-DN-NFYA (lane 2), Mef after 48 hours post infection with Ad-DN-NFYA (lane 3). The extracts were probed with rabbit polyclonal with anti-NFY-A. To normalised protein's loading, the filter was stained with a monoclonal antibody anti-actin protein. (C) Q-ChIP analysis, of the indicated NF-Y target promoters on MEF after 16, 24, 48 hours (h) from Ad-GFP (white bars), or Ad-DN-NFYA after 16 h from infection (dark grey bars), 24 h (black bars), 48 h (grey bars), performed with anti-NFY-A, -p300, -H4K20me3, -H3K9me3 and -H3K9ac antibodies. The fold difference value in each case compares the sample performed after Ad-DN-NFYA infection to the corresponding control sample performed after Ad-GFP infection at the same time point (defined as 1). One of two independent experiments performed in triplicate is represented. (D) Q-ChIP analysis, performed with anti-HP1α antibody, of the indicated NF-Y target promoters on MEF after 24 h after infection with Ad-GFP (white bars) and Ad-DN-NFYA (black bars). It is shown one of two independent experiments performed in triplicate. (E) ChIPs were performed on proliferating MEFs 24 hours after Ad-GFP and wtNF-YA infections. No antibody was used as control (No Ab). PCR analysis were performed on the immunoprecipitated DNA samples with anti-NFY-A, -H3K9ac and -PAN-H4ac-Pan antibodies, using specific primers for the indicate promoters.
    Figure Legend Snippet: Lack of NF-Y DNA binding activity induces hypermethylation of histones and target gene inactivation. (A) RT-PCR amplification of the indicated genes from uninfected MEFs (lane 1), MEFs 24hours after infection with Ad-GFP (lane 2), MEFs 24hours and 48hours after infection with Ad-DN-NFYA (lanes 3 and 4). (B) Western blot analysis performed on total lysates from: Mef after 24hours post infection with Ad-GFP (lane1), Mef after 24 hours post infection with Ad-DN-NFYA (lane 2), Mef after 48 hours post infection with Ad-DN-NFYA (lane 3). The extracts were probed with rabbit polyclonal with anti-NFY-A. To normalised protein's loading, the filter was stained with a monoclonal antibody anti-actin protein. (C) Q-ChIP analysis, of the indicated NF-Y target promoters on MEF after 16, 24, 48 hours (h) from Ad-GFP (white bars), or Ad-DN-NFYA after 16 h from infection (dark grey bars), 24 h (black bars), 48 h (grey bars), performed with anti-NFY-A, -p300, -H4K20me3, -H3K9me3 and -H3K9ac antibodies. The fold difference value in each case compares the sample performed after Ad-DN-NFYA infection to the corresponding control sample performed after Ad-GFP infection at the same time point (defined as 1). One of two independent experiments performed in triplicate is represented. (D) Q-ChIP analysis, performed with anti-HP1α antibody, of the indicated NF-Y target promoters on MEF after 24 h after infection with Ad-GFP (white bars) and Ad-DN-NFYA (black bars). It is shown one of two independent experiments performed in triplicate. (E) ChIPs were performed on proliferating MEFs 24 hours after Ad-GFP and wtNF-YA infections. No antibody was used as control (No Ab). PCR analysis were performed on the immunoprecipitated DNA samples with anti-NFY-A, -H3K9ac and -PAN-H4ac-Pan antibodies, using specific primers for the indicate promoters.

    Techniques Used: Binding Assay, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Amplification, Infection, Western Blot, Staining, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Immunoprecipitation

    6) Product Images from "UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery"

    Article Title: UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery

    Journal: Genes & Development

    doi: 10.1101/gad.310705

    Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined
    Figure Legend Snippet: Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined

    Techniques Used: Clone Assay

    7) Product Images from "UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery"

    Article Title: UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery

    Journal: Genes & Development

    doi: 10.1101/gad.310705

    Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined
    Figure Legend Snippet: Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined

    Techniques Used: Clone Assay

    8) Product Images from "Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation"

    Article Title: Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007532

    Transcription analysis of cells where VSG synthesis is blocked. A) Northern blot analysis of T. brucei 221VG1.1 cells where VSG221 RNAi had been induced for the time indicated above in hours (h). The parental (P) T. brucei 90-13 cell line does not contain the VSG221 RNAi construct. The T. brucei 221VG1.1 cell line was either not incubated with tetracycline (− Tet) or had VSG221 RNAi induced with tetracycline for the times indicated above. Blots were hybridised with probes for Actin, NUP1 , eGFP (present in the active VSG221 ES), TDP1 , tubulin (Tub), ESAG5 , 18S rRNA, ISWI , ESAG6/7, VSG221 and PFR2 . Ethidium stains of the gels are indicated on the right to indicate total RNA loaded. B) Transcription analysis of T. brucei 221VG1.1 where VSG synthesis was blocked by the induction of VSG221 RNAi with tetracycline (Tet) for 0 or 24 hours (h). Cells were incubated with BrUTP to label nascent transcripts and subsequently incubated with an anti-BrdUTP antibody, and a secondary antibody coupled to Alexa 488. DNA was stained with DAPI. A normal precytokinesis cell (0 h) is compared with a precytokinesis cell arising after the induction of VSG RNAi for 24 hours (24 h). The experiment was performed in the absence of α-amanitin (− α-ama) to visualise total transcription, or in the presence of 200 µg ml −1 α-amanitin to inhibit transcription by RNA polymerases II and III and visualise transcription by RNA polymerase I (+ α-ama). The scale bar indicates 4 µm. Quantitation of transcription as fluorescence in the FITC channel is in arbitrary units using 50 cells per time point, with standard deviation indicated with error bars.
    Figure Legend Snippet: Transcription analysis of cells where VSG synthesis is blocked. A) Northern blot analysis of T. brucei 221VG1.1 cells where VSG221 RNAi had been induced for the time indicated above in hours (h). The parental (P) T. brucei 90-13 cell line does not contain the VSG221 RNAi construct. The T. brucei 221VG1.1 cell line was either not incubated with tetracycline (− Tet) or had VSG221 RNAi induced with tetracycline for the times indicated above. Blots were hybridised with probes for Actin, NUP1 , eGFP (present in the active VSG221 ES), TDP1 , tubulin (Tub), ESAG5 , 18S rRNA, ISWI , ESAG6/7, VSG221 and PFR2 . Ethidium stains of the gels are indicated on the right to indicate total RNA loaded. B) Transcription analysis of T. brucei 221VG1.1 where VSG synthesis was blocked by the induction of VSG221 RNAi with tetracycline (Tet) for 0 or 24 hours (h). Cells were incubated with BrUTP to label nascent transcripts and subsequently incubated with an anti-BrdUTP antibody, and a secondary antibody coupled to Alexa 488. DNA was stained with DAPI. A normal precytokinesis cell (0 h) is compared with a precytokinesis cell arising after the induction of VSG RNAi for 24 hours (24 h). The experiment was performed in the absence of α-amanitin (− α-ama) to visualise total transcription, or in the presence of 200 µg ml −1 α-amanitin to inhibit transcription by RNA polymerases II and III and visualise transcription by RNA polymerase I (+ α-ama). The scale bar indicates 4 µm. Quantitation of transcription as fluorescence in the FITC channel is in arbitrary units using 50 cells per time point, with standard deviation indicated with error bars.

    Techniques Used: Northern Blot, Construct, Incubation, Staining, Quantitation Assay, Fluorescence, Standard Deviation

    9) Product Images from "Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation"

    Article Title: Blocking Variant Surface Glycoprotein Synthesis in Trypanosoma brucei Triggers a General Arrest in Translation Initiation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007532

    Transcription analysis of cells where VSG synthesis is blocked. A) Northern blot analysis of T. brucei 221VG1.1 cells where VSG221 RNAi had been induced for the time indicated above in hours (h). The parental (P) T. brucei 90-13 cell line does not contain the VSG221 RNAi construct. The T. brucei 221VG1.1 cell line was either not incubated with tetracycline (− Tet) or had VSG221 RNAi induced with tetracycline for the times indicated above. Blots were hybridised with probes for Actin, NUP1 , eGFP (present in the active VSG221 ES), TDP1 , tubulin (Tub), ESAG5 , 18S rRNA, ISWI , ESAG6/7, VSG221 and PFR2 . Ethidium stains of the gels are indicated on the right to indicate total RNA loaded. B) Transcription analysis of T. brucei 221VG1.1 where VSG synthesis was blocked by the induction of VSG221 RNAi with tetracycline (Tet) for 0 or 24 hours (h). Cells were incubated with BrUTP to label nascent transcripts and subsequently incubated with an anti-BrdUTP antibody, and a secondary antibody coupled to Alexa 488. DNA was stained with DAPI. A normal precytokinesis cell (0 h) is compared with a precytokinesis cell arising after the induction of VSG RNAi for 24 hours (24 h). The experiment was performed in the absence of α-amanitin (− α-ama) to visualise total transcription, or in the presence of 200 µg ml −1 α-amanitin to inhibit transcription by RNA polymerases II and III and visualise transcription by RNA polymerase I (+ α-ama). The scale bar indicates 4 µm. Quantitation of transcription as fluorescence in the FITC channel is in arbitrary units using 50 cells per time point, with standard deviation indicated with error bars.
    Figure Legend Snippet: Transcription analysis of cells where VSG synthesis is blocked. A) Northern blot analysis of T. brucei 221VG1.1 cells where VSG221 RNAi had been induced for the time indicated above in hours (h). The parental (P) T. brucei 90-13 cell line does not contain the VSG221 RNAi construct. The T. brucei 221VG1.1 cell line was either not incubated with tetracycline (− Tet) or had VSG221 RNAi induced with tetracycline for the times indicated above. Blots were hybridised with probes for Actin, NUP1 , eGFP (present in the active VSG221 ES), TDP1 , tubulin (Tub), ESAG5 , 18S rRNA, ISWI , ESAG6/7, VSG221 and PFR2 . Ethidium stains of the gels are indicated on the right to indicate total RNA loaded. B) Transcription analysis of T. brucei 221VG1.1 where VSG synthesis was blocked by the induction of VSG221 RNAi with tetracycline (Tet) for 0 or 24 hours (h). Cells were incubated with BrUTP to label nascent transcripts and subsequently incubated with an anti-BrdUTP antibody, and a secondary antibody coupled to Alexa 488. DNA was stained with DAPI. A normal precytokinesis cell (0 h) is compared with a precytokinesis cell arising after the induction of VSG RNAi for 24 hours (24 h). The experiment was performed in the absence of α-amanitin (− α-ama) to visualise total transcription, or in the presence of 200 µg ml −1 α-amanitin to inhibit transcription by RNA polymerases II and III and visualise transcription by RNA polymerase I (+ α-ama). The scale bar indicates 4 µm. Quantitation of transcription as fluorescence in the FITC channel is in arbitrary units using 50 cells per time point, with standard deviation indicated with error bars.

    Techniques Used: Northern Blot, Construct, Incubation, Staining, Quantitation Assay, Fluorescence, Standard Deviation

    10) Product Images from "UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery"

    Article Title: UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery

    Journal: Genes & Development

    doi: 10.1101/gad.310705

    Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined
    Figure Legend Snippet: Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined

    Techniques Used: Clone Assay

    11) Product Images from "UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery"

    Article Title: UBF-binding site arrays form pseudo-NORs and sequester the RNA polymerase I transcription machinery

    Journal: Genes & Development

    doi: 10.1101/gad.310705

    Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined
    Figure Legend Snippet: Pseudo-NORs are transcriptionally inactive. Ongoing pol I-mediated transcription was visualized in clones 5A, 4E, 5B, and 5E by incorporation of BrUTP into nascent transcripts. ( Left panels) BrUTP was visualized with anti-BrdUTP antibodies (Roche) combined

    Techniques Used: Clone Assay

    Related Articles

    Centrifugation:

    Article Title: Human Cytomegalovirus Glycoprotein B Is Required for Virus Entry and Cell-to-Cell Spread but Not for Virion Attachment, Assembly, or Egress ▿
    Article Snippet: .. Six days postinfection, infected cell supernatant was collected, cellular debris was removed by centrifugation, and for real-time PCR, any contaminating DNA was removed by DNase I (Roche Applied Science, Indianapolis, IN) digestion at 37°C for 30 min. DNA from the supernatant was extracted and analyzed to detect viral genomes by real-time PCR as described above. .. To purify released virions, the infected cell supernatant was concentrated by pelleting it over a 20% sorbitol cushion, followed by banding in a 20 to 70% sorbitol step gradient.

    WST-1 Assay:

    Article Title: The Ubiquitin-Proteasome System Plays an Important Role during Various Stages of the Coronavirus Infection Cycle ▿
    Article Snippet: .. At 6 h postinfection, the cell number and viability was measured by Wst-1 assay according to the manufacturer's protocol (Roche Diagnostics GmbH). .. Subsequently, intracellular luciferase expression was determined as described above.

    Protease Inhibitor:

    Article Title: Antagonism of dsRNA-Induced Innate Immune Pathways by NS4a and NS4b Accessory Proteins during MERS Coronavirus Infection
    Article Snippet: .. Cells were washed twice with ice-cold PBS and lysates harvested at indicated times postinfection with lysis buffer (1% NP-40, 2 mM EDTA, 10% glycerol, 150 mM NaCl, 50 mM Tris HCl) supplemented with protease inhibitors (Roche cOmplete mini-EDTA-free protease inhibitor) and phosphatase inhibitors (Roche PhosStop Easy Pack). .. Samples were heated at 95°C for 5 min and then se parated by 4 to 15% SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes.

    Transfection:

    Article Title: The impact of intragenic CpG content on gene expression
    Article Snippet: .. One-hour postinfection, cells were washed and transfected with pS/huGFP60 and pS/huGFP0 constructs, respectively, using FuGENE 6 (Roche). .. Cells were cultured for another 12 h, and protein expression was analysed by fluorescent-activated cell sorting (FACS) as described.

    Infection:

    Article Title: Human Cytomegalovirus Glycoprotein B Is Required for Virus Entry and Cell-to-Cell Spread but Not for Virion Attachment, Assembly, or Egress ▿
    Article Snippet: .. Six days postinfection, infected cell supernatant was collected, cellular debris was removed by centrifugation, and for real-time PCR, any contaminating DNA was removed by DNase I (Roche Applied Science, Indianapolis, IN) digestion at 37°C for 30 min. DNA from the supernatant was extracted and analyzed to detect viral genomes by real-time PCR as described above. .. To purify released virions, the infected cell supernatant was concentrated by pelleting it over a 20% sorbitol cushion, followed by banding in a 20 to 70% sorbitol step gradient.

    Real-time Polymerase Chain Reaction:

    Article Title: Human Cytomegalovirus Glycoprotein B Is Required for Virus Entry and Cell-to-Cell Spread but Not for Virion Attachment, Assembly, or Egress ▿
    Article Snippet: .. Six days postinfection, infected cell supernatant was collected, cellular debris was removed by centrifugation, and for real-time PCR, any contaminating DNA was removed by DNase I (Roche Applied Science, Indianapolis, IN) digestion at 37°C for 30 min. DNA from the supernatant was extracted and analyzed to detect viral genomes by real-time PCR as described above. .. To purify released virions, the infected cell supernatant was concentrated by pelleting it over a 20% sorbitol cushion, followed by banding in a 20 to 70% sorbitol step gradient.

    Construct:

    Article Title: The impact of intragenic CpG content on gene expression
    Article Snippet: .. One-hour postinfection, cells were washed and transfected with pS/huGFP60 and pS/huGFP0 constructs, respectively, using FuGENE 6 (Roche). .. Cells were cultured for another 12 h, and protein expression was analysed by fluorescent-activated cell sorting (FACS) as described.

    Cotransfection:

    Article Title: RNA-Templated Replication of Hepatitis Delta Virus: Genomic and Antigenomic RNAs Associate with Different Nuclear Bodies
    Article Snippet: .. For BrUTP and HDV RNA cotransfection, DOTAP (Roche) was used. ..

    Staining:

    Article Title: The g5R (D250) Gene of African Swine Fever Virus Encodes a Nudix Hydrolase That Preferentially Degrades Diphosphoinositol Polyphosphates
    Article Snippet: .. At various times postinfection (7, 12, and 16 h) cells were fixed in 4% paraformaldehyde, permeabilized in 0.2% Triton X-100, and stained with rat anti-HA monoclonal antibody (1 in 600 dilution; Roche) and Alexa Fluor 568 goat anti-rat immunoglobulin G conjugate (1 in 800 dilution; Molecular Probes). .. Cells were visualized with a Leica TCS NT confocal microscope.

    Lysis:

    Article Title: Antagonism of dsRNA-Induced Innate Immune Pathways by NS4a and NS4b Accessory Proteins during MERS Coronavirus Infection
    Article Snippet: .. Cells were washed twice with ice-cold PBS and lysates harvested at indicated times postinfection with lysis buffer (1% NP-40, 2 mM EDTA, 10% glycerol, 150 mM NaCl, 50 mM Tris HCl) supplemented with protease inhibitors (Roche cOmplete mini-EDTA-free protease inhibitor) and phosphatase inhibitors (Roche PhosStop Easy Pack). .. Samples were heated at 95°C for 5 min and then se parated by 4 to 15% SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes.

    Article Title: Flavivirus Infection Impairs Peroxisome Biogenesis and Early Antiviral Signaling
    Article Snippet: .. At designated time points posttransfection or postinfection, the cells were washed three times with phosphate-buffered saline (PBS) before lysing with NP-40 lysis buffer (150 mM NaCl, 2 mM EDTA, 1% Nonidet P-40, 50 mM Tris-HCl [pH 7.4], 1 mM dithiothreitol) containing Complete protease inhibitors (Roche) on ice for 30 min. .. The lysates were clarified at 14,000 rpm for 20 min in a microcentrifuge at 4°C and precleared with protein G or protein A Sepharose beads for 5 min at 4°C, followed by incubation with appropriate antibodies for several hours at 4°C.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Roche anti brutp
    (A) NF-Y is associated with active transcription sites in living cells. After in vivo incorporation of <t>BrUTP</t> (run-on), cells were fixed and endogenous NF-YA (ii) and nascent RNA transcripts (i) were detected by indirect immunofluorescence combined with Confocal Scanning Laser Microscopy by using anti-NF-YA and anti-BrU antibodies. In the overlay (iii), yellow indicates colocalizations between NF-YA (green) and transcription sites (red). In panels vi and x cells were immunostained with anti-NF-YA, in panels v and ix with <t>anti-RPII</t> CTD repeat YSPTSPS (phospho S2) and anti-total RPII respectively. The majority of NF-YA (red) colocalizes with the activated form of RPII (green)(vii). (B) Cells were immunostained with anti-NF-YA (vii-xii), -acetylated H3K9 (i), -acetylated H4 (ii), -tri-methylated H3K9 (iii), -di-methylated H3K27 (iv), -tri-methylated H4K20 (v) and -NF-YB (vi) antibodies. The majority of NF-YA colocalizes with acetylated (xiii, xiv), but poor colocalization occurs with methylated histones (xv, xvi,xvii). Panel xviii shows the overlay of two subunits of NF-Y, NF-YA (xii) and NF-YB (vi). Panels from xix to xxiv represent a typical optical field of the merge. In figure 1A and 1B confocal analysis of single optical section is shown. The images have been collected with a 60x objective.
    Anti Brutp, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti brutp/product/Roche
    Average 85 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    anti brutp - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    Image Search Results


    (A) NF-Y is associated with active transcription sites in living cells. After in vivo incorporation of BrUTP (run-on), cells were fixed and endogenous NF-YA (ii) and nascent RNA transcripts (i) were detected by indirect immunofluorescence combined with Confocal Scanning Laser Microscopy by using anti-NF-YA and anti-BrU antibodies. In the overlay (iii), yellow indicates colocalizations between NF-YA (green) and transcription sites (red). In panels vi and x cells were immunostained with anti-NF-YA, in panels v and ix with anti-RPII CTD repeat YSPTSPS (phospho S2) and anti-total RPII respectively. The majority of NF-YA (red) colocalizes with the activated form of RPII (green)(vii). (B) Cells were immunostained with anti-NF-YA (vii-xii), -acetylated H3K9 (i), -acetylated H4 (ii), -tri-methylated H3K9 (iii), -di-methylated H3K27 (iv), -tri-methylated H4K20 (v) and -NF-YB (vi) antibodies. The majority of NF-YA colocalizes with acetylated (xiii, xiv), but poor colocalization occurs with methylated histones (xv, xvi,xvii). Panel xviii shows the overlay of two subunits of NF-Y, NF-YA (xii) and NF-YB (vi). Panels from xix to xxiv represent a typical optical field of the merge. In figure 1A and 1B confocal analysis of single optical section is shown. The images have been collected with a 60x objective.

    Journal: PLoS ONE

    Article Title: NF-Y Dependent Epigenetic Modifications Discriminate between Proliferating and Postmitotic Tissue

    doi: 10.1371/journal.pone.0002047

    Figure Lengend Snippet: (A) NF-Y is associated with active transcription sites in living cells. After in vivo incorporation of BrUTP (run-on), cells were fixed and endogenous NF-YA (ii) and nascent RNA transcripts (i) were detected by indirect immunofluorescence combined with Confocal Scanning Laser Microscopy by using anti-NF-YA and anti-BrU antibodies. In the overlay (iii), yellow indicates colocalizations between NF-YA (green) and transcription sites (red). In panels vi and x cells were immunostained with anti-NF-YA, in panels v and ix with anti-RPII CTD repeat YSPTSPS (phospho S2) and anti-total RPII respectively. The majority of NF-YA (red) colocalizes with the activated form of RPII (green)(vii). (B) Cells were immunostained with anti-NF-YA (vii-xii), -acetylated H3K9 (i), -acetylated H4 (ii), -tri-methylated H3K9 (iii), -di-methylated H3K27 (iv), -tri-methylated H4K20 (v) and -NF-YB (vi) antibodies. The majority of NF-YA colocalizes with acetylated (xiii, xiv), but poor colocalization occurs with methylated histones (xv, xvi,xvii). Panel xviii shows the overlay of two subunits of NF-Y, NF-YA (xii) and NF-YB (vi). Panels from xix to xxiv represent a typical optical field of the merge. In figure 1A and 1B confocal analysis of single optical section is shown. The images have been collected with a 60x objective.

    Article Snippet: The following primary antibodies (diluted in 1% BSA) were used: anti-NF-YA rabbit polyclonal (on run on experiments) (Rockland n°200-401-100, Gilbertsville, PA), anti-NF-YA mouse monoclonal from hybridoma cells, anti-NF-YB rabbit polyclonal (gifts from R. Mantovani), anti-BrUTP (Roche Diagnostics n°1170376, clone BMC 9318), anti-RPII (Santa Cruz n° 899), anti- RPIIS2 (Abcam n°5095), anti-PAN-H4ac (Upstate n°06-598), anti-H3ac-k9 (Abcam n°4441), anti-H3tri-m-k9 (Abcam n°8898), anti-H4tri-m-k20 (Abcam n°9053), anti-p300 (Santa Cruz n° 584 and 585), anti-H3di-m-k27 (Abcam n°24684) and anti-HDAC1 (Sigma H3284).

    Techniques: In Vivo, Immunofluorescence, Microscopy, Methylation