immunoprecipitation  (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
  • 86

    Structured Review

    Roche immunoprecipitation
    Immunoprecipitation, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitation - by Bioz Stars, 2021-07
    86/100 stars

    Images

    Related Articles

    Co-Immunoprecipitation Assay:

    Article Title: Suppression of 14-3-3γ-mediated surface expression of ANO1 inhibits cancer progression of glioblastoma cells
    Article Snippet: To assess the protein-protein interaction between ANO1 and 14-3-3γ, BD/ANO1 seg (a ) and AD/14-3-3γ were co-transformed into the yeast strain AH109, which was incubated in selection medium. .. Co-IP and immunoblotting For Co-IP, the lysates of transfected HEK293T cells and U251 cells were prepared with lysis buffer (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 5 mM EDTA, and 0.1% Triton X-100) containing a protease-inhibitor cocktail (Roche) and mixed overnight at 4 °C with anti-HA (F-7; Santa Cruz Biotechnology), anti-FLAG (M-2; Sigma), or anti-ANO1 (ab53212; Abcam) antibodies. .. Immune complexes were incubated by binding to mixed protein A/G beads (Santa Cruz Biotechnology) for 1 h. For immunoblotting, protein samples were separated by 10% SDS-PAGE gel under the same experimental condition and then transferred to PVDF membranes.

    Transfection:

    Article Title: Suppression of 14-3-3γ-mediated surface expression of ANO1 inhibits cancer progression of glioblastoma cells
    Article Snippet: To assess the protein-protein interaction between ANO1 and 14-3-3γ, BD/ANO1 seg (a ) and AD/14-3-3γ were co-transformed into the yeast strain AH109, which was incubated in selection medium. .. Co-IP and immunoblotting For Co-IP, the lysates of transfected HEK293T cells and U251 cells were prepared with lysis buffer (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 5 mM EDTA, and 0.1% Triton X-100) containing a protease-inhibitor cocktail (Roche) and mixed overnight at 4 °C with anti-HA (F-7; Santa Cruz Biotechnology), anti-FLAG (M-2; Sigma), or anti-ANO1 (ab53212; Abcam) antibodies. .. Immune complexes were incubated by binding to mixed protein A/G beads (Santa Cruz Biotechnology) for 1 h. For immunoblotting, protein samples were separated by 10% SDS-PAGE gel under the same experimental condition and then transferred to PVDF membranes.

    Article Title: Erk regulation of actin capping and bundling by Eps8 promotes cortex tension and leader bleb-based migration
    Article Snippet: Bands were then resolved on an Odyssey scanner (LI-COR Biosciences). .. Immunoprecipitations 12 hr after transfection of plasmids encoding EGFP or Emerald-tagged mouse Eps8, A375 whole-cell lysates were prepared by scrapeing into ice cold RIPA buffer (50 mM Hepes pH 7.4, 150 mM NaCl, 5 mM EDTA, 0.1% SDS, 0.5% deoxycholate and 1% Triton X-100) containing protease and phosphatase inhibitors (Roche). ..

    Article Title: TRAPPC9 Mediates the Interaction between p150Glued and COPII Vesicles at the Target Membrane
    Article Snippet: Transfection assay was performed with polyethylenimine (PEI) method (Godbey et al., 1999) or PolyJet (SL100688) according to the manufacturer's instructions (SignaGene Lab). .. Immunoprecipitation Transfected COS1 cells in 10 cm tissue culture plates were lysed in lysis buffer (20 mM Tris, pH 8.0, 100 mM NaCl, 0.1% NP-40 and 1× protease inhibitor cocktail Complete® (Roche)). .. The supernatants were subject to immunoprecipitation with 1–2 µg of anti-Myc or anti-FLAG antibody and 30 µl of protein-A sepharose slurry (sigma, P3391).

    Article Title: Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿ †
    Article Snippet: After extensive washing, horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) were applied, and the membrane was washed again and then developed with a chemiluminescence reagent (Perkin-Elmer, Waltham, MA) and visualized on a Kodak Image Station 4000R (Kodak Molecular Imaging Systems, Rochester, NY). .. Transfected 293T cells were lysed in high-salt NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 40 mM Tris-HCl [pH 7.5], 300 mM NaCl, and 10 mM MgCl2 ) containing EDTA-free Complete protease inhibitor (Roche, Mannheim, Germany) and cleared by centrifugation at 10,000 × g for 10 min. Supernatants were diluted 1:1 with 1% (vol/vol) NP-40 and precleared with Sepharose (Sigma) for 2 h at 4°C before incubation with anti-Flag M2 agarose (Sigma) overnight at 4°C. .. The beads were washed extensively with NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 20 mM Tris-HCl [pH 7.5], 150 mM NaCl, and 5 mM MgCl2 ) and eluted with 0.4 mg/ml Flag peptide (Sigma) in NP-40 lysis buffer.

    Lysis:

    Article Title: Suppression of 14-3-3γ-mediated surface expression of ANO1 inhibits cancer progression of glioblastoma cells
    Article Snippet: To assess the protein-protein interaction between ANO1 and 14-3-3γ, BD/ANO1 seg (a ) and AD/14-3-3γ were co-transformed into the yeast strain AH109, which was incubated in selection medium. .. Co-IP and immunoblotting For Co-IP, the lysates of transfected HEK293T cells and U251 cells were prepared with lysis buffer (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 5 mM EDTA, and 0.1% Triton X-100) containing a protease-inhibitor cocktail (Roche) and mixed overnight at 4 °C with anti-HA (F-7; Santa Cruz Biotechnology), anti-FLAG (M-2; Sigma), or anti-ANO1 (ab53212; Abcam) antibodies. .. Immune complexes were incubated by binding to mixed protein A/G beads (Santa Cruz Biotechnology) for 1 h. For immunoblotting, protein samples were separated by 10% SDS-PAGE gel under the same experimental condition and then transferred to PVDF membranes.

    Article Title: Characterization of Mammalian Selenoprotein O: A Redox-Active Mitochondrial Protein
    Article Snippet: .. 75 Se Metabolic Labeling and Identifying Human SelO HEK293T cells were metabolically labeled with 75 Se for 40 h. The cells were lysed in 0.1% NP40 cell lysis buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 5% Glycerol) containing protease inhibitor (Roche Applied Science, Indianapolis, IN, USA) and 20 mM N -ethylmaleimide. .. The cell lysates were sonicated for 15 min and centrifuged at 13,000 rpm for 10 min at 4°C.

    Article Title: TRAPPC9 Mediates the Interaction between p150Glued and COPII Vesicles at the Target Membrane
    Article Snippet: Transfection assay was performed with polyethylenimine (PEI) method (Godbey et al., 1999) or PolyJet (SL100688) according to the manufacturer's instructions (SignaGene Lab). .. Immunoprecipitation Transfected COS1 cells in 10 cm tissue culture plates were lysed in lysis buffer (20 mM Tris, pH 8.0, 100 mM NaCl, 0.1% NP-40 and 1× protease inhibitor cocktail Complete® (Roche)). .. The supernatants were subject to immunoprecipitation with 1–2 µg of anti-Myc or anti-FLAG antibody and 30 µl of protein-A sepharose slurry (sigma, P3391).

    Article Title: Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿ †
    Article Snippet: After extensive washing, horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) were applied, and the membrane was washed again and then developed with a chemiluminescence reagent (Perkin-Elmer, Waltham, MA) and visualized on a Kodak Image Station 4000R (Kodak Molecular Imaging Systems, Rochester, NY). .. Transfected 293T cells were lysed in high-salt NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 40 mM Tris-HCl [pH 7.5], 300 mM NaCl, and 10 mM MgCl2 ) containing EDTA-free Complete protease inhibitor (Roche, Mannheim, Germany) and cleared by centrifugation at 10,000 × g for 10 min. Supernatants were diluted 1:1 with 1% (vol/vol) NP-40 and precleared with Sepharose (Sigma) for 2 h at 4°C before incubation with anti-Flag M2 agarose (Sigma) overnight at 4°C. .. The beads were washed extensively with NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 20 mM Tris-HCl [pH 7.5], 150 mM NaCl, and 5 mM MgCl2 ) and eluted with 0.4 mg/ml Flag peptide (Sigma) in NP-40 lysis buffer.

    Protease Inhibitor:

    Article Title: Suppression of 14-3-3γ-mediated surface expression of ANO1 inhibits cancer progression of glioblastoma cells
    Article Snippet: To assess the protein-protein interaction between ANO1 and 14-3-3γ, BD/ANO1 seg (a ) and AD/14-3-3γ were co-transformed into the yeast strain AH109, which was incubated in selection medium. .. Co-IP and immunoblotting For Co-IP, the lysates of transfected HEK293T cells and U251 cells were prepared with lysis buffer (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 5 mM EDTA, and 0.1% Triton X-100) containing a protease-inhibitor cocktail (Roche) and mixed overnight at 4 °C with anti-HA (F-7; Santa Cruz Biotechnology), anti-FLAG (M-2; Sigma), or anti-ANO1 (ab53212; Abcam) antibodies. .. Immune complexes were incubated by binding to mixed protein A/G beads (Santa Cruz Biotechnology) for 1 h. For immunoblotting, protein samples were separated by 10% SDS-PAGE gel under the same experimental condition and then transferred to PVDF membranes.

    Article Title: Characterization of Mammalian Selenoprotein O: A Redox-Active Mitochondrial Protein
    Article Snippet: .. 75 Se Metabolic Labeling and Identifying Human SelO HEK293T cells were metabolically labeled with 75 Se for 40 h. The cells were lysed in 0.1% NP40 cell lysis buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 5% Glycerol) containing protease inhibitor (Roche Applied Science, Indianapolis, IN, USA) and 20 mM N -ethylmaleimide. .. The cell lysates were sonicated for 15 min and centrifuged at 13,000 rpm for 10 min at 4°C.

    Article Title: TRAPPC9 Mediates the Interaction between p150Glued and COPII Vesicles at the Target Membrane
    Article Snippet: Transfection assay was performed with polyethylenimine (PEI) method (Godbey et al., 1999) or PolyJet (SL100688) according to the manufacturer's instructions (SignaGene Lab). .. Immunoprecipitation Transfected COS1 cells in 10 cm tissue culture plates were lysed in lysis buffer (20 mM Tris, pH 8.0, 100 mM NaCl, 0.1% NP-40 and 1× protease inhibitor cocktail Complete® (Roche)). .. The supernatants were subject to immunoprecipitation with 1–2 µg of anti-Myc or anti-FLAG antibody and 30 µl of protein-A sepharose slurry (sigma, P3391).

    Article Title: Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿ †
    Article Snippet: After extensive washing, horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) were applied, and the membrane was washed again and then developed with a chemiluminescence reagent (Perkin-Elmer, Waltham, MA) and visualized on a Kodak Image Station 4000R (Kodak Molecular Imaging Systems, Rochester, NY). .. Transfected 293T cells were lysed in high-salt NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 40 mM Tris-HCl [pH 7.5], 300 mM NaCl, and 10 mM MgCl2 ) containing EDTA-free Complete protease inhibitor (Roche, Mannheim, Germany) and cleared by centrifugation at 10,000 × g for 10 min. Supernatants were diluted 1:1 with 1% (vol/vol) NP-40 and precleared with Sepharose (Sigma) for 2 h at 4°C before incubation with anti-Flag M2 agarose (Sigma) overnight at 4°C. .. The beads were washed extensively with NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 20 mM Tris-HCl [pH 7.5], 150 mM NaCl, and 5 mM MgCl2 ) and eluted with 0.4 mg/ml Flag peptide (Sigma) in NP-40 lysis buffer.

    Article Title: Lucanthone and Its Derivative Hycanthone Inhibit Apurinic Endonuclease-1 (APE1) by Direct Protein Binding
    Article Snippet: The endonuclease activity inhibition by lucanthone was analyzed using standard Lineweaver-Burke Plot to determine if the inhibition was competitive or non-competitive. .. Cleavage of APE1 by lucanthone and CRT0044876 Western blotting was carried out by 7.5% SDS-PAGE of cell extracts (20 µg total protein per lane) either from APE1 overexpresser clone 5 pretreated with 2.5–200 µM concentration of lucanthone/CRT0044876 or recombinant APE1 for 2 h at 37°C in presence of protease inhibitor cocktail (2 tablets (Roche, # 11836153001) containing mixture of several protease inhibitors with broad inhibitory specificity for serine, cysteine and metalloproteases in all systems, dissolved in 20 ml of APE1 buffer). ..

    Article Title: Rab27a controls HIV-1 assembly by regulating plasma membrane levels of phosphatidylinositol 4,5-bisphosphate
    Article Snippet: The supernatant contained the isolated membrane proteins. .. Immunoprecipitation of GFP-CD63 For immunoprecipitation of GFP-CD63, 107 GFP-CD63–transfected cells were lysed in immunoprecipitation buffer (1% Brij 99, 10 mM Tris/HCl, pH 7.4, 150 mM NaCl, and 5 mM EDTA, with Roche protease inhibitor cocktail) for 1 h at 4°C. .. Precleared supernatants were then immunoprecipitated with GFP-Traps or uncoated agarose beads (as control for unspecific bound) according to manufacturer’s instructions (ChromoTek).

    Polymerase Chain Reaction:

    Article Title: Regulation of DNA replication at the end of the mitochondrial D-loop involves the helicase TWINKLE and a conserved sequence element
    Article Snippet: PCR products were separated by agarose gel electrophoresis and cloned with the Zero blunt TOPO PCR Cloning Kit (Invitogen) before sequencing. .. Quantification of mtDNA levels Total DNA was extracted with High Pure PCR Template Preparation Kit (Roche). ..

    Labeling:

    Article Title: Characterization of Mammalian Selenoprotein O: A Redox-Active Mitochondrial Protein
    Article Snippet: .. 75 Se Metabolic Labeling and Identifying Human SelO HEK293T cells were metabolically labeled with 75 Se for 40 h. The cells were lysed in 0.1% NP40 cell lysis buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 5% Glycerol) containing protease inhibitor (Roche Applied Science, Indianapolis, IN, USA) and 20 mM N -ethylmaleimide. .. The cell lysates were sonicated for 15 min and centrifuged at 13,000 rpm for 10 min at 4°C.

    Metabolic Labelling:

    Article Title: Characterization of Mammalian Selenoprotein O: A Redox-Active Mitochondrial Protein
    Article Snippet: .. 75 Se Metabolic Labeling and Identifying Human SelO HEK293T cells were metabolically labeled with 75 Se for 40 h. The cells were lysed in 0.1% NP40 cell lysis buffer (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 5% Glycerol) containing protease inhibitor (Roche Applied Science, Indianapolis, IN, USA) and 20 mM N -ethylmaleimide. .. The cell lysates were sonicated for 15 min and centrifuged at 13,000 rpm for 10 min at 4°C.

    Immunoprecipitation:

    Article Title: TRAPPC9 Mediates the Interaction between p150Glued and COPII Vesicles at the Target Membrane
    Article Snippet: Transfection assay was performed with polyethylenimine (PEI) method (Godbey et al., 1999) or PolyJet (SL100688) according to the manufacturer's instructions (SignaGene Lab). .. Immunoprecipitation Transfected COS1 cells in 10 cm tissue culture plates were lysed in lysis buffer (20 mM Tris, pH 8.0, 100 mM NaCl, 0.1% NP-40 and 1× protease inhibitor cocktail Complete® (Roche)). .. The supernatants were subject to immunoprecipitation with 1–2 µg of anti-Myc or anti-FLAG antibody and 30 µl of protein-A sepharose slurry (sigma, P3391).

    Article Title: Rab27a controls HIV-1 assembly by regulating plasma membrane levels of phosphatidylinositol 4,5-bisphosphate
    Article Snippet: The supernatant contained the isolated membrane proteins. .. Immunoprecipitation of GFP-CD63 For immunoprecipitation of GFP-CD63, 107 GFP-CD63–transfected cells were lysed in immunoprecipitation buffer (1% Brij 99, 10 mM Tris/HCl, pH 7.4, 150 mM NaCl, and 5 mM EDTA, with Roche protease inhibitor cocktail) for 1 h at 4°C. .. Precleared supernatants were then immunoprecipitated with GFP-Traps or uncoated agarose beads (as control for unspecific bound) according to manufacturer’s instructions (ChromoTek).

    Centrifugation:

    Article Title: Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿ †
    Article Snippet: After extensive washing, horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) were applied, and the membrane was washed again and then developed with a chemiluminescence reagent (Perkin-Elmer, Waltham, MA) and visualized on a Kodak Image Station 4000R (Kodak Molecular Imaging Systems, Rochester, NY). .. Transfected 293T cells were lysed in high-salt NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 40 mM Tris-HCl [pH 7.5], 300 mM NaCl, and 10 mM MgCl2 ) containing EDTA-free Complete protease inhibitor (Roche, Mannheim, Germany) and cleared by centrifugation at 10,000 × g for 10 min. Supernatants were diluted 1:1 with 1% (vol/vol) NP-40 and precleared with Sepharose (Sigma) for 2 h at 4°C before incubation with anti-Flag M2 agarose (Sigma) overnight at 4°C. .. The beads were washed extensively with NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 20 mM Tris-HCl [pH 7.5], 150 mM NaCl, and 5 mM MgCl2 ) and eluted with 0.4 mg/ml Flag peptide (Sigma) in NP-40 lysis buffer.

    Incubation:

    Article Title: Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿Epstein-Barr Virus LF2 Protein Regulates Viral Replication by Altering Rta Subcellular Localization ▿ †
    Article Snippet: After extensive washing, horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) were applied, and the membrane was washed again and then developed with a chemiluminescence reagent (Perkin-Elmer, Waltham, MA) and visualized on a Kodak Image Station 4000R (Kodak Molecular Imaging Systems, Rochester, NY). .. Transfected 293T cells were lysed in high-salt NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 40 mM Tris-HCl [pH 7.5], 300 mM NaCl, and 10 mM MgCl2 ) containing EDTA-free Complete protease inhibitor (Roche, Mannheim, Germany) and cleared by centrifugation at 10,000 × g for 10 min. Supernatants were diluted 1:1 with 1% (vol/vol) NP-40 and precleared with Sepharose (Sigma) for 2 h at 4°C before incubation with anti-Flag M2 agarose (Sigma) overnight at 4°C. .. The beads were washed extensively with NP-40 lysis buffer (1% [vol/vol] Igepal CA-630, 20 mM Tris-HCl [pH 7.5], 150 mM NaCl, and 5 mM MgCl2 ) and eluted with 0.4 mg/ml Flag peptide (Sigma) in NP-40 lysis buffer.

    Western Blot:

    Article Title: Lucanthone and Its Derivative Hycanthone Inhibit Apurinic Endonuclease-1 (APE1) by Direct Protein Binding
    Article Snippet: The endonuclease activity inhibition by lucanthone was analyzed using standard Lineweaver-Burke Plot to determine if the inhibition was competitive or non-competitive. .. Cleavage of APE1 by lucanthone and CRT0044876 Western blotting was carried out by 7.5% SDS-PAGE of cell extracts (20 µg total protein per lane) either from APE1 overexpresser clone 5 pretreated with 2.5–200 µM concentration of lucanthone/CRT0044876 or recombinant APE1 for 2 h at 37°C in presence of protease inhibitor cocktail (2 tablets (Roche, # 11836153001) containing mixture of several protease inhibitors with broad inhibitory specificity for serine, cysteine and metalloproteases in all systems, dissolved in 20 ml of APE1 buffer). ..

    SDS Page:

    Article Title: Lucanthone and Its Derivative Hycanthone Inhibit Apurinic Endonuclease-1 (APE1) by Direct Protein Binding
    Article Snippet: The endonuclease activity inhibition by lucanthone was analyzed using standard Lineweaver-Burke Plot to determine if the inhibition was competitive or non-competitive. .. Cleavage of APE1 by lucanthone and CRT0044876 Western blotting was carried out by 7.5% SDS-PAGE of cell extracts (20 µg total protein per lane) either from APE1 overexpresser clone 5 pretreated with 2.5–200 µM concentration of lucanthone/CRT0044876 or recombinant APE1 for 2 h at 37°C in presence of protease inhibitor cocktail (2 tablets (Roche, # 11836153001) containing mixture of several protease inhibitors with broad inhibitory specificity for serine, cysteine and metalloproteases in all systems, dissolved in 20 ml of APE1 buffer). ..

    Concentration Assay:

    Article Title: Lucanthone and Its Derivative Hycanthone Inhibit Apurinic Endonuclease-1 (APE1) by Direct Protein Binding
    Article Snippet: The endonuclease activity inhibition by lucanthone was analyzed using standard Lineweaver-Burke Plot to determine if the inhibition was competitive or non-competitive. .. Cleavage of APE1 by lucanthone and CRT0044876 Western blotting was carried out by 7.5% SDS-PAGE of cell extracts (20 µg total protein per lane) either from APE1 overexpresser clone 5 pretreated with 2.5–200 µM concentration of lucanthone/CRT0044876 or recombinant APE1 for 2 h at 37°C in presence of protease inhibitor cocktail (2 tablets (Roche, # 11836153001) containing mixture of several protease inhibitors with broad inhibitory specificity for serine, cysteine and metalloproteases in all systems, dissolved in 20 ml of APE1 buffer). ..

    Recombinant:

    Article Title: Lucanthone and Its Derivative Hycanthone Inhibit Apurinic Endonuclease-1 (APE1) by Direct Protein Binding
    Article Snippet: The endonuclease activity inhibition by lucanthone was analyzed using standard Lineweaver-Burke Plot to determine if the inhibition was competitive or non-competitive. .. Cleavage of APE1 by lucanthone and CRT0044876 Western blotting was carried out by 7.5% SDS-PAGE of cell extracts (20 µg total protein per lane) either from APE1 overexpresser clone 5 pretreated with 2.5–200 µM concentration of lucanthone/CRT0044876 or recombinant APE1 for 2 h at 37°C in presence of protease inhibitor cocktail (2 tablets (Roche, # 11836153001) containing mixture of several protease inhibitors with broad inhibitory specificity for serine, cysteine and metalloproteases in all systems, dissolved in 20 ml of APE1 buffer). ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    Roche immunoprecipitation cells
    EGFR-MET interaction is modulated by EGFR mutations. (A) Western blot (WB) of total MET levels in H1975 derived cells. Tubulin levels are shown as loading control. Values beneath blots are relative levels of MET compared to the total levels in the H1975 L858R/T790M cell line from 2 independent experiments +/−SD (B) MET (7q31) (red signal) copy number analysis by FISH in the three H1975 cell lines using the Leica Kreatech C-MET (7q31)/SE7 FISH probe (KBI-10719). The green signal indicates the chromosome 7 centromere control probe. Scale bar 10 mm. Average copy number and ratio between MET and chromosome 7 centromere probe are also indicated (n = 30 cells). (C) Immunofluorescence of total EGFR (Alexa546 –red in the image) and MET (Cyanine 5 –green in the image) in H1975 derived cells. Hoescht dye was used to stain the nuclei of the cells. Merge panels are also shown. Bars, 20 μm. (D) <t>Co-immunoprecipitation</t> (IP) of EGFR in H1975 derived cell lines. The EGFR antibody was used to immunoprecipitate. EGFR and MET levels are shown in both bound and input fractions. The gels shown in the figure were run separately for the bound and input fractions, as indicated by the dotted line, under the same experimental conditions. (E) Fluorescence lifetime imaging was performed on cells plated to sub-confluence on cover-slips and time-resolved analysis in Tri2. Quantification of average FRET efficiency (*** p
    Immunoprecipitation Cells, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation cells/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitation cells - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    86
    Roche immunoprecipitations
    Figure 6. Large-scale screen for phosphorylated LC3 interaction partners. ( A ) Using a rat eGFP-MAP1LC3-MCF7 cell line <t>immunoprecipitations</t> by anti-GFP antibodies were performed to enrich LC3-interacting proteins from untreated control cells and
    Immunoprecipitations, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitations/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitations - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    86
    Roche immunoprecipitation buffer
    NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of <t>immunoprecipitation</t> eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.
    Immunoprecipitation Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation buffer/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitation buffer - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    Image Search Results


    EGFR-MET interaction is modulated by EGFR mutations. (A) Western blot (WB) of total MET levels in H1975 derived cells. Tubulin levels are shown as loading control. Values beneath blots are relative levels of MET compared to the total levels in the H1975 L858R/T790M cell line from 2 independent experiments +/−SD (B) MET (7q31) (red signal) copy number analysis by FISH in the three H1975 cell lines using the Leica Kreatech C-MET (7q31)/SE7 FISH probe (KBI-10719). The green signal indicates the chromosome 7 centromere control probe. Scale bar 10 mm. Average copy number and ratio between MET and chromosome 7 centromere probe are also indicated (n = 30 cells). (C) Immunofluorescence of total EGFR (Alexa546 –red in the image) and MET (Cyanine 5 –green in the image) in H1975 derived cells. Hoescht dye was used to stain the nuclei of the cells. Merge panels are also shown. Bars, 20 μm. (D) Co-immunoprecipitation (IP) of EGFR in H1975 derived cell lines. The EGFR antibody was used to immunoprecipitate. EGFR and MET levels are shown in both bound and input fractions. The gels shown in the figure were run separately for the bound and input fractions, as indicated by the dotted line, under the same experimental conditions. (E) Fluorescence lifetime imaging was performed on cells plated to sub-confluence on cover-slips and time-resolved analysis in Tri2. Quantification of average FRET efficiency (*** p

    Journal: PLoS ONE

    Article Title: MET-EGFR dimerization in lung adenocarcinoma is dependent on EGFR mtations and altered by MET kinase inhibition

    doi: 10.1371/journal.pone.0170798

    Figure Lengend Snippet: EGFR-MET interaction is modulated by EGFR mutations. (A) Western blot (WB) of total MET levels in H1975 derived cells. Tubulin levels are shown as loading control. Values beneath blots are relative levels of MET compared to the total levels in the H1975 L858R/T790M cell line from 2 independent experiments +/−SD (B) MET (7q31) (red signal) copy number analysis by FISH in the three H1975 cell lines using the Leica Kreatech C-MET (7q31)/SE7 FISH probe (KBI-10719). The green signal indicates the chromosome 7 centromere control probe. Scale bar 10 mm. Average copy number and ratio between MET and chromosome 7 centromere probe are also indicated (n = 30 cells). (C) Immunofluorescence of total EGFR (Alexa546 –red in the image) and MET (Cyanine 5 –green in the image) in H1975 derived cells. Hoescht dye was used to stain the nuclei of the cells. Merge panels are also shown. Bars, 20 μm. (D) Co-immunoprecipitation (IP) of EGFR in H1975 derived cell lines. The EGFR antibody was used to immunoprecipitate. EGFR and MET levels are shown in both bound and input fractions. The gels shown in the figure were run separately for the bound and input fractions, as indicated by the dotted line, under the same experimental conditions. (E) Fluorescence lifetime imaging was performed on cells plated to sub-confluence on cover-slips and time-resolved analysis in Tri2. Quantification of average FRET efficiency (*** p

    Article Snippet: Immunoprecipitation Cells were lysed in lysis buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 10% glycerol, 1% Triton X-100, 10 mM NaF, 1 mM Na3 VO4 , 10 mM N-ethylmaleimide, 0.01 μM Calyculin A) with Protease inhibitor cocktail set I (Roche).

    Techniques: Western Blot, Derivative Assay, Fluorescence In Situ Hybridization, Immunofluorescence, Staining, Immunoprecipitation, Fluorescence, Imaging

    Figure 6. Large-scale screen for phosphorylated LC3 interaction partners. ( A ) Using a rat eGFP-MAP1LC3-MCF7 cell line immunoprecipitations by anti-GFP antibodies were performed to enrich LC3-interacting proteins from untreated control cells and

    Journal: Autophagy

    Article Title: Characterization of early autophagy signaling by quantitative phosphoproteomics

    doi: 10.4161/auto.26864

    Figure Lengend Snippet: Figure 6. Large-scale screen for phosphorylated LC3 interaction partners. ( A ) Using a rat eGFP-MAP1LC3-MCF7 cell line immunoprecipitations by anti-GFP antibodies were performed to enrich LC3-interacting proteins from untreated control cells and

    Article Snippet: For autophagosome accumulation cells were treated with 2 nM conA (Sigma-Aldrich, 27689) for 7 h. Anti-GFP (Santa Cruz Biotechnology, sc-9996) immunoprecipitations of SILAC-labeled MCF7 cells expressing rat eGFP-MAP1LC3 were performed in modified RIPA buffer containing 1% NP-40, 150 mM NaCl, 0.25% Na deoxycholate, 50 mM Tris pH 7.5 and Complete Protease Inhibitor tablets (Roche, 11836145001).

    Techniques:

    In vitro G β binding specificity of GGL domain mutants. ( A ) Secondary structure predictions for the RGS6 GGL domain and sequence alignment between G γ 2 ) are plotted above the primary sequence of the GGL domain ( x axis). α-Helices within G γ 2 ) are indicated by an α above the sequence. The position and nature of point mutations are denoted above or below the sequence line with arrows. Individual G β subunits were cotranslated in reticulocyte lysates with wild-type or mutant RGS6 ( B ), RGS7 ( C ), and RGS11 proteins ( D–F ). HA-tagged RGS or G γ proteins were immunoprecipitated in low detergent (except as noted in E ) with anti-HA mAb, and associated G β proteins were visualized by SDS/PAGE and autoradiography. ( E ) Immunoprecipitations (IP) of cotranslated G β 5 and wild-type (lane 1) or W274F mutant (lanes 2 and 3) RGS11ΔDΔC proteins were performed in high-detergent (lanes 1 and 3) or low-detergent (lane 2) conditions and visualized separately from clarified supernatants (Sup’nt) as above.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Fidelity of G protein ?-subunit association by the G protein ?-subunit-like domains of RGS6, RGS7, and RGS11

    doi:

    Figure Lengend Snippet: In vitro G β binding specificity of GGL domain mutants. ( A ) Secondary structure predictions for the RGS6 GGL domain and sequence alignment between G γ 2 ) are plotted above the primary sequence of the GGL domain ( x axis). α-Helices within G γ 2 ) are indicated by an α above the sequence. The position and nature of point mutations are denoted above or below the sequence line with arrows. Individual G β subunits were cotranslated in reticulocyte lysates with wild-type or mutant RGS6 ( B ), RGS7 ( C ), and RGS11 proteins ( D–F ). HA-tagged RGS or G γ proteins were immunoprecipitated in low detergent (except as noted in E ) with anti-HA mAb, and associated G β proteins were visualized by SDS/PAGE and autoradiography. ( E ) Immunoprecipitations (IP) of cotranslated G β 5 and wild-type (lane 1) or W274F mutant (lanes 2 and 3) RGS11ΔDΔC proteins were performed in high-detergent (lanes 1 and 3) or low-detergent (lane 2) conditions and visualized separately from clarified supernatants (Sup’nt) as above.

    Article Snippet: “Low-detergent” immunoprecipitations were performed and washed in buffer D [50 mM NaCl/10 mM MgCl2 /50 mM Tris, pH 8.0/1 mM EDTA/10 mM 2-mercaptoethanol/20% (vol/vol) glycerol/Complete protease inhibitors; Roche Diagnostics] containing 0.05% C12E10, whereas “high-detergent” immunoprecipitations were performed in buffer D containing 0.1% Triton X-100 and washed in RIPA-500 buffer containing 500 mM NaCl, 1% Triton X-100, 0.5% deoxycholate, and 0.1% SDS.

    Techniques: In Vitro, Binding Assay, Sequencing, Mutagenesis, Immunoprecipitation, SDS Page, Autoradiography

    NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Journal: The Journal of Neuroscience

    Article Title: Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity

    doi: 10.1523/JNEUROSCI.2548-14.2015

    Figure Lengend Snippet: NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Article Snippet: For total rat brain extraction, the whole rat brain was homogenized in 10 volumes of immunoprecipitation buffer (50 m m Tris-HCl, pH 7.5, 100 m m NaCl, 2 m m CaCl2 , 1% Triton X-100) containing the mini-EDTA-free protease inhibitor cocktail (Roche).

    Techniques: Western Blot, Immunoprecipitation, Molecular Weight, Recombinant, Transfection, Immunofluorescence, Confocal Microscopy, Isolation, Negative Control, Marker, Cell Culture