biotin conjugated goat anti rabbit igg Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Vector Laboratories biotin conjugated goat anti rabbit igg
    Expression of <t>PI3Kγ</t> protein in mouse ASM cells. A, representative immunohistochemical staining of PI3Kγ protein in mouse trachea (a) and lung slices (c) using a PI3Kγ-specific antibody or isotype-specific negative control antibody (rabbit <t>IgG)</t> in a parallel section of trachea (b) and lung slices (d). PI3Kγ staining was localized in ASM and epithelium but not in cartilage. All the sections were counterstained with blue hematoxylin. B, Western blot analysis of PI3Kγ protein in lysates of mouse lung tissue, heart tissue, and isolated mouse ASM cells using a PI3Kγ-specific antibody. Images shown are representative of four independent experiments. β-Actin was used as a loading control. C, immunofluorescent staining of PI3Kγ protein (green) in cultured mouse ASM cells, and staining of α-actin (red) as a marker of smooth muscle cells.
    Biotin Conjugated Goat Anti Rabbit Igg, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 973 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotin conjugated goat anti rabbit igg/product/Vector Laboratories
    Average 99 stars, based on 973 article reviews
    Price from $9.99 to $1999.99
    biotin conjugated goat anti rabbit igg - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Millipore anti rabbit igg whole molecule peroxidase antibody
    Expression of <t>PI3Kγ</t> protein in mouse ASM cells. A, representative immunohistochemical staining of PI3Kγ protein in mouse trachea (a) and lung slices (c) using a PI3Kγ-specific antibody or isotype-specific negative control antibody (rabbit <t>IgG)</t> in a parallel section of trachea (b) and lung slices (d). PI3Kγ staining was localized in ASM and epithelium but not in cartilage. All the sections were counterstained with blue hematoxylin. B, Western blot analysis of PI3Kγ protein in lysates of mouse lung tissue, heart tissue, and isolated mouse ASM cells using a PI3Kγ-specific antibody. Images shown are representative of four independent experiments. β-Actin was used as a loading control. C, immunofluorescent staining of PI3Kγ protein (green) in cultured mouse ASM cells, and staining of α-actin (red) as a marker of smooth muscle cells.
    Anti Rabbit Igg Whole Molecule Peroxidase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7032 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit igg whole molecule peroxidase antibody/product/Millipore
    Average 99 stars, based on 7032 article reviews
    Price from $9.99 to $1999.99
    anti rabbit igg whole molecule peroxidase antibody - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Rockland Immunochemicals goat anti rabbit
    Interaction of N782 with the XRCC5/6 complex. * Indicates EGFP tag. ( a ) Left panel shows proteins immunoprecipitated with a rabbit <t>anti-GFP</t> antibody (black circle) and co-immunoprecipitated proteins (arrow). It contains both rabbit <t>IgG</t> heavy chain (50 kD wide band) and the light chain (25 kD wide band) derived from the anti-GFP antibody. Right panel shows Western blots of immunoprecipitated EGFP-KDM2A, EGFP-N782, and EGFP-KDM2A C-termini. The proteins were detected by a combination of mouse anti-GFP (primary) and goat anti-mouse IgG. ( b ) Keratinocyte genomic DNA fragments co-immunoprecipitated with EGFP-N782 and EGFP. A marker (250 or 500 ng) or sample (5 μl) was applied to each well, respectively. ( c ) Mass spectrometry of proteins included in the 70–90 kDa protein bands [proteins from top to bottom, (XRCC5 and XRCC6 in red) and (TMPO and LMNA in blue)]. ( d ) Western blotting analysis of XRCC5 (arrow) co-immunoprecipitated with N782 or the mutants (circle): Δ = deletion, C = CXXC, J = JmjC, and P = PHD. ( e ) Binding of GST-CN, GST-PN, and GST-NLSR to 3-length IGFBPL1 baits. Full-length blots are presented in Supplementary Fig. S1 . ( f ) Far Western blotting analysis of the interaction of KMD2A domains with XRCC5 and 6. Probe and protein dye are on the left; cross-reaction of antibody to an E. coli protein, > and
    Goat Anti Rabbit, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 99/100, based on 278 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit/product/Rockland Immunochemicals
    Average 99 stars, based on 278 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Bio-Rad hrp conjugated goat anti rabbit igg
    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with <t>HRP-conjugated</t> anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat <t>IgG</t> and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 485 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti rabbit igg/product/Bio-Rad
    Average 99 stars, based on 485 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated goat anti rabbit igg - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Boster Bio goat anti rabbit igg h l secondary antibody biotin conjugate
    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with <t>HRP-conjugated</t> anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat <t>IgG</t> and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.
    Goat Anti Rabbit Igg H L Secondary Antibody Biotin Conjugate, supplied by Boster Bio, used in various techniques. Bioz Stars score: 99/100, based on 340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg h l secondary antibody biotin conjugate/product/Boster Bio
    Average 99 stars, based on 340 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg h l secondary antibody biotin conjugate - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Bio-Rad goat anti rabbit igg hrp conjugate
    Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit <t>IgG</t> antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). ( a ) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; ( b ) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance ( p
    Goat Anti Rabbit Igg Hrp Conjugate, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 365 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg hrp conjugate/product/Bio-Rad
    Average 99 stars, based on 365 article reviews
    Price from $9.99 to $1999.99
    goat anti rabbit igg hrp conjugate - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Agilent technologies biotin conjugated goat anti rabbit igg
    Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit <t>IgG</t> antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). ( a ) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; ( b ) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance ( p
    Biotin Conjugated Goat Anti Rabbit Igg, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotin conjugated goat anti rabbit igg/product/Agilent technologies
    Average 92 stars, based on 104 article reviews
    Price from $9.99 to $1999.99
    biotin conjugated goat anti rabbit igg - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    99
    Boster Bio hrp conjugated anti rabbit igg sabc kit
    Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit <t>IgG</t> antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). ( a ) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; ( b ) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance ( p
    Hrp Conjugated Anti Rabbit Igg Sabc Kit, supplied by Boster Bio, used in various techniques. Bioz Stars score: 99/100, based on 561 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated anti rabbit igg sabc kit/product/Boster Bio
    Average 99 stars, based on 561 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated anti rabbit igg sabc kit - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Jackson Immuno biotin sp affinipure goat anti rabbit igg
    Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase <t>(HRP)-conjugated</t> goat anti-rabbit <t>IgG</t> antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). ( a ) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; ( b ) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance ( p
    Biotin Sp Affinipure Goat Anti Rabbit Igg, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 99/100, based on 574 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotin sp affinipure goat anti rabbit igg/product/Jackson Immuno
    Average 99 stars, based on 574 article reviews
    Price from $9.99 to $1999.99
    biotin sp affinipure goat anti rabbit igg - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Thermo Fisher biotin conjugated goat anti rabbit igg
    Proximal tubule cells secrete <t>pro-AOAH.</t> (A) Diagram of AOAH structure, showing the conversion of the precursor (pro-AOAH) into mature AOAH. (B) Production of 35 S-AOAH by porcine proximal tubule cells in vitro. Labeled AOAH was immunoprecipitated as described in Methods and studied by SDS-PAGE and autoradiography. M, medium; L, lysate. Antibody +, anti-AOAH; antibody −, preimmune <t>IgG.</t> Only pro-AOAH is seen in the media, whereas the lysate contains both pro-AOAH and mature AOAH.
    Biotin Conjugated Goat Anti Rabbit Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotin conjugated goat anti rabbit igg/product/Thermo Fisher
    Average 92 stars, based on 69 article reviews
    Price from $9.99 to $1999.99
    biotin conjugated goat anti rabbit igg - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    95
    Millipore anti rabbit igg whole molecule biotin antibody
    Proximal tubule cells secrete <t>pro-AOAH.</t> (A) Diagram of AOAH structure, showing the conversion of the precursor (pro-AOAH) into mature AOAH. (B) Production of 35 S-AOAH by porcine proximal tubule cells in vitro. Labeled AOAH was immunoprecipitated as described in Methods and studied by SDS-PAGE and autoradiography. M, medium; L, lysate. Antibody +, anti-AOAH; antibody −, preimmune <t>IgG.</t> Only pro-AOAH is seen in the media, whereas the lysate contains both pro-AOAH and mature AOAH.
    Anti Rabbit Igg Whole Molecule Biotin Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 341 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit igg whole molecule biotin antibody/product/Millipore
    Average 95 stars, based on 341 article reviews
    Price from $9.99 to $1999.99
    anti rabbit igg whole molecule biotin antibody - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    Image Search Results


    Expression of PI3Kγ protein in mouse ASM cells. A, representative immunohistochemical staining of PI3Kγ protein in mouse trachea (a) and lung slices (c) using a PI3Kγ-specific antibody or isotype-specific negative control antibody (rabbit IgG) in a parallel section of trachea (b) and lung slices (d). PI3Kγ staining was localized in ASM and epithelium but not in cartilage. All the sections were counterstained with blue hematoxylin. B, Western blot analysis of PI3Kγ protein in lysates of mouse lung tissue, heart tissue, and isolated mouse ASM cells using a PI3Kγ-specific antibody. Images shown are representative of four independent experiments. β-Actin was used as a loading control. C, immunofluorescent staining of PI3Kγ protein (green) in cultured mouse ASM cells, and staining of α-actin (red) as a marker of smooth muscle cells.

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Phosphoinositide 3-Kinase ? Regulates Airway Smooth Muscle Contraction by Modulating Calcium Oscillations

    doi: 10.1124/jpet.110.168518

    Figure Lengend Snippet: Expression of PI3Kγ protein in mouse ASM cells. A, representative immunohistochemical staining of PI3Kγ protein in mouse trachea (a) and lung slices (c) using a PI3Kγ-specific antibody or isotype-specific negative control antibody (rabbit IgG) in a parallel section of trachea (b) and lung slices (d). PI3Kγ staining was localized in ASM and epithelium but not in cartilage. All the sections were counterstained with blue hematoxylin. B, Western blot analysis of PI3Kγ protein in lysates of mouse lung tissue, heart tissue, and isolated mouse ASM cells using a PI3Kγ-specific antibody. Images shown are representative of four independent experiments. β-Actin was used as a loading control. C, immunofluorescent staining of PI3Kγ protein (green) in cultured mouse ASM cells, and staining of α-actin (red) as a marker of smooth muscle cells.

    Article Snippet: Rabbit PI3Kγ antibody and biotin-conjugated goat anti-rabbit IgG from Vector Laboratories (Burlingame, CA) were used as the primary antibody and secondary antibody, respectively.

    Techniques: Expressing, Immunohistochemistry, Staining, Negative Control, Western Blot, Isolation, Cell Culture, Marker

    Interaction of N782 with the XRCC5/6 complex. * Indicates EGFP tag. ( a ) Left panel shows proteins immunoprecipitated with a rabbit anti-GFP antibody (black circle) and co-immunoprecipitated proteins (arrow). It contains both rabbit IgG heavy chain (50 kD wide band) and the light chain (25 kD wide band) derived from the anti-GFP antibody. Right panel shows Western blots of immunoprecipitated EGFP-KDM2A, EGFP-N782, and EGFP-KDM2A C-termini. The proteins were detected by a combination of mouse anti-GFP (primary) and goat anti-mouse IgG. ( b ) Keratinocyte genomic DNA fragments co-immunoprecipitated with EGFP-N782 and EGFP. A marker (250 or 500 ng) or sample (5 μl) was applied to each well, respectively. ( c ) Mass spectrometry of proteins included in the 70–90 kDa protein bands [proteins from top to bottom, (XRCC5 and XRCC6 in red) and (TMPO and LMNA in blue)]. ( d ) Western blotting analysis of XRCC5 (arrow) co-immunoprecipitated with N782 or the mutants (circle): Δ = deletion, C = CXXC, J = JmjC, and P = PHD. ( e ) Binding of GST-CN, GST-PN, and GST-NLSR to 3-length IGFBPL1 baits. Full-length blots are presented in Supplementary Fig. S1 . ( f ) Far Western blotting analysis of the interaction of KMD2A domains with XRCC5 and 6. Probe and protein dye are on the left; cross-reaction of antibody to an E. coli protein, > and

    Journal: Scientific Reports

    Article Title: Lysine-specific demethylase 2A enhances binding of various nuclear factors to CpG-rich genomic DNAs by action of its CXXC-PHD domain

    doi: 10.1038/s41598-019-41896-6

    Figure Lengend Snippet: Interaction of N782 with the XRCC5/6 complex. * Indicates EGFP tag. ( a ) Left panel shows proteins immunoprecipitated with a rabbit anti-GFP antibody (black circle) and co-immunoprecipitated proteins (arrow). It contains both rabbit IgG heavy chain (50 kD wide band) and the light chain (25 kD wide band) derived from the anti-GFP antibody. Right panel shows Western blots of immunoprecipitated EGFP-KDM2A, EGFP-N782, and EGFP-KDM2A C-termini. The proteins were detected by a combination of mouse anti-GFP (primary) and goat anti-mouse IgG. ( b ) Keratinocyte genomic DNA fragments co-immunoprecipitated with EGFP-N782 and EGFP. A marker (250 or 500 ng) or sample (5 μl) was applied to each well, respectively. ( c ) Mass spectrometry of proteins included in the 70–90 kDa protein bands [proteins from top to bottom, (XRCC5 and XRCC6 in red) and (TMPO and LMNA in blue)]. ( d ) Western blotting analysis of XRCC5 (arrow) co-immunoprecipitated with N782 or the mutants (circle): Δ = deletion, C = CXXC, J = JmjC, and P = PHD. ( e ) Binding of GST-CN, GST-PN, and GST-NLSR to 3-length IGFBPL1 baits. Full-length blots are presented in Supplementary Fig. S1 . ( f ) Far Western blotting analysis of the interaction of KMD2A domains with XRCC5 and 6. Probe and protein dye are on the left; cross-reaction of antibody to an E. coli protein, > and

    Article Snippet: The following antibodies were used for Western blotting: rabbit anti-ORC3 (Thermofisher, PA5-28083); rabbit anti-POLR1A (Thermofisher, PA5-37156); mouse anti-XRCC5 (Thermo Scientific, MA5-15873); mouse anti-XRCC6 (Abnova, H00002547-M01), rabbit polyclonal anti-H3 (Thermofisher Scientific, PA-16183), mouse anti-H2B (Biolegend, 688702), anti-GFP monoclonal antibody (Clontech), rabbit anti-GFP (Acris, SP3005P); goat anti-rabbit IgG (H + L), IRDy 800 conjugated (Rockland, 611-132-122); and IRDye 680RD goat anti-mouse IgG (H + L) (LI-COR, 926-68170).

    Techniques: Immunoprecipitation, Derivative Assay, Western Blot, Marker, Mass Spectrometry, Binding Assay, Far Western Blot

    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with HRP-conjugated anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat IgG and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.

    Journal: The Journal of Biological Chemistry

    Article Title: New Insights into the DT40 B Cell Receptor Cluster Using a Proteomic Proximity Labeling Assay *

    doi: 10.1074/jbc.M113.529578

    Figure Lengend Snippet: SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with HRP-conjugated anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat IgG and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.

    Article Snippet: About 5 × 108 cells were incubated with end over end rotation for 2 h with 20 μg/ml of HRP-conjugated goat anti-chicken IgM (Bethyl Ltd.), or 20 μg/ml of HRP-conjugated goat anti-rabbit IgG (Bio-Rad) in 20 ml of PBS with 10% (v/v) goat serum (Ultraclone Ltd.) at 4 °C.

    Techniques: Labeling, Avidin-Biotin Assay, Staining

    Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). ( a ) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; ( b ) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance ( p

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: The Antiproliferative Effect of Cyclodipeptides from Pseudomonas aeruginosa PAO1 on HeLa Cells Involves Inhibition of Phosphorylation of Akt and S6k Kinases

    doi: 10.3390/molecules22061024

    Figure Lengend Snippet: Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). ( a ) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; ( b ) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance ( p

    Article Snippet: After washing, the membrane was incubated with the secondary antibody, Goat anti-Rabbit IgG HRP-conjugate (BioRad), in blocking medium at 1:10,000 dilution for 4 h at 4 °C; the membrane was twice washed with TBS-T and developed using hydrogen peroxide and Supersignal West Pico Luminol (Pierce, Thermo Fisher Scientific) and after exposing in light-sensitive films or ChemiDoc™ MP System (Bio-Rad).

    Techniques: Expressing, Incubation, Sonication, Polyacrylamide Gel Electrophoresis, SDS Page, Software, Immunodetection, Quantitation Assay

    Proximal tubule cells secrete pro-AOAH. (A) Diagram of AOAH structure, showing the conversion of the precursor (pro-AOAH) into mature AOAH. (B) Production of 35 S-AOAH by porcine proximal tubule cells in vitro. Labeled AOAH was immunoprecipitated as described in Methods and studied by SDS-PAGE and autoradiography. M, medium; L, lysate. Antibody +, anti-AOAH; antibody −, preimmune IgG. Only pro-AOAH is seen in the media, whereas the lysate contains both pro-AOAH and mature AOAH.

    Journal: Infection and Immunity

    Article Title: Identification of Acyloxyacyl Hydrolase, a Lipopolysaccharide- Detoxifying Enzyme, in the Murine Urinary Tract

    doi: 10.1128/IAI.72.6.3171-3178.2004

    Figure Lengend Snippet: Proximal tubule cells secrete pro-AOAH. (A) Diagram of AOAH structure, showing the conversion of the precursor (pro-AOAH) into mature AOAH. (B) Production of 35 S-AOAH by porcine proximal tubule cells in vitro. Labeled AOAH was immunoprecipitated as described in Methods and studied by SDS-PAGE and autoradiography. M, medium; L, lysate. Antibody +, anti-AOAH; antibody −, preimmune IgG. Only pro-AOAH is seen in the media, whereas the lysate contains both pro-AOAH and mature AOAH.

    Article Snippet: Wells were blocked for 1 h at 37°C, samples and recombinant human AOAH standard were incubated at 4°C overnight or at 37°C for 2 h, secondary antibody (E553, purified polyclonal rabbit anti-human AOAH, final concentration 10 μg/ml) was added for 1 h at 37°C, biotin-conjugated goat anti-rabbit IgG (0.075 μg/ml) (Zymed, San Francisco, Calif.) was added for 1 h at 37°C, 2 μg of alkaline phosphatase-conjugated streptavidin (Jackson Laboratories)/ml was incubated for 30 min at 37°C, and the plate was developed with alkaline phosphatase substrate (5 mM Sigma 104 and 0.1 M Sigma alkaline buffer 221 in water).

    Techniques: In Vitro, Labeling, Immunoprecipitation, SDS Page, Autoradiography