nonidet p 40  (Santa Cruz Biotechnology)

 
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    Name:
    Nonidet P40
    Description:
    Nonidet P40 substitute is a non ionic non denaturing detergent suitable for solubilizing and isolation of membrane proteins Has been used to solubilize cerebral GABA receptors Also useful in solubilization of proteins and lipids of the scallop gill ciliary membrane and of membranes from breast cancer tumors Chemically indistinguishable from Nonidet P 40 which is no longer commercially available
    Catalog Number:
    SC-29102
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    Chemicals Other Chemicals Polymers Nonidet P40 substitute
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    Nonidet® P40 (substitute) is a non-ionic, non-denaturing detergent primarily used to solubilize membrane proteins
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    Structured Review

    Santa Cruz Biotechnology nonidet p 40
    PGC-1α interacts with OGT. A , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and OGT co-immunoprecipitation ( IP ). Immunoprecipitates were  subjected to SDS-PAGE and immunoblotted ( IB ) for the presence of  PGC-1α. Membranes were then stripped and blotted for OGT. Immunoblots  are representative of three experiments.  B , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and FLAG co-immunoprecipitation. Immunoprecipitates were subjected to SDS-PAGE  and immunoblotted for the presence of OGT. Membranes were then stripped and  blotted for PGC-1α.  C , gel filtration chromatography (SMART  system, Superdex 200 column, phosphate-buffered saline, 1% Nonidet P-40  buffer) of lysates from rat liver incubated on ice for 30 min with either  normal IgG or anti-OGT antibodies (AL28). Fractions were subjected to SDS-PAGE  and blotted using anti-PGC-1α, anti-OGT (DM-17), and anti CPB. Data are  representative of two experiments.
    Nonidet P40 substitute is a non ionic non denaturing detergent suitable for solubilizing and isolation of membrane proteins Has been used to solubilize cerebral GABA receptors Also useful in solubilization of proteins and lipids of the scallop gill ciliary membrane and of membranes from breast cancer tumors Chemically indistinguishable from Nonidet P 40 which is no longer commercially available
    https://www.bioz.com/result/nonidet p 40/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nonidet p 40 - by Bioz Stars, 2021-05
    93/100 stars

    Images

    1) Product Images from "A PGC-1α-O-GlcNAc Transferase Complex Regulates FoxO Transcription Factor Activity in Response to Glucose"

    Article Title: A PGC-1α-O-GlcNAc Transferase Complex Regulates FoxO Transcription Factor Activity in Response to Glucose

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M808890200

    PGC-1α interacts with OGT. A , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and OGT co-immunoprecipitation ( IP ). Immunoprecipitates were  subjected to SDS-PAGE and immunoblotted ( IB ) for the presence of  PGC-1α. Membranes were then stripped and blotted for OGT. Immunoblots  are representative of three experiments.  B , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and FLAG co-immunoprecipitation. Immunoprecipitates were subjected to SDS-PAGE  and immunoblotted for the presence of OGT. Membranes were then stripped and  blotted for PGC-1α.  C , gel filtration chromatography (SMART  system, Superdex 200 column, phosphate-buffered saline, 1% Nonidet P-40  buffer) of lysates from rat liver incubated on ice for 30 min with either  normal IgG or anti-OGT antibodies (AL28). Fractions were subjected to SDS-PAGE  and blotted using anti-PGC-1α, anti-OGT (DM-17), and anti CPB. Data are  representative of two experiments.
    Figure Legend Snippet: PGC-1α interacts with OGT. A , Fao cells were infected with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting and OGT co-immunoprecipitation ( IP ). Immunoprecipitates were subjected to SDS-PAGE and immunoblotted ( IB ) for the presence of PGC-1α. Membranes were then stripped and blotted for OGT. Immunoblots are representative of three experiments. B , Fao cells were infected with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting and FLAG co-immunoprecipitation. Immunoprecipitates were subjected to SDS-PAGE and immunoblotted for the presence of OGT. Membranes were then stripped and blotted for PGC-1α. C , gel filtration chromatography (SMART system, Superdex 200 column, phosphate-buffered saline, 1% Nonidet P-40 buffer) of lysates from rat liver incubated on ice for 30 min with either normal IgG or anti-OGT antibodies (AL28). Fractions were subjected to SDS-PAGE and blotted using anti-PGC-1α, anti-OGT (DM-17), and anti CPB. Data are representative of two experiments.

    Techniques Used: Pyrolysis Gas Chromatography, Infection, Immunoprecipitation, SDS Page, Western Blot, Filtration, Chromatography, Incubation

    2) Product Images from "Covalent modification of the homeodomain-interacting protein kinase 2 (HIPK2) by the ubiquitin-like protein SUMO-1"

    Article Title: Covalent modification of the homeodomain-interacting protein kinase 2 (HIPK2) by the ubiquitin-like protein SUMO-1

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

    doi:

    Localization of SUMO-1-modified HIPK2 to nuclear speckles (dots). ( A ) Fractionation of SUMO-1-modified HIPK2. Cellular extracts from CV-1 cells cotransfected with GFP-SUMO-1 and Myc-HIPK2 expression vectors were fractionated. Proteins from each fraction were separated on SDS/polyacrylamide gradient (4–12%) gels and analyzed by Western blotting using a mouse anti-Myc Ab. HIPK2 and SUMO-1-modified HIPK2 are detected in the nuclear fraction (lane 3) and in the Nonidet P-40-insoluble fractions (lanes 7 and 8). SUMO-1-modified HIPK2 and unmodified HIPK2 are marked by bracket and arrowhead, respectively. ( B ) Confocal microscopic image of the nucleus showing colocalization of HIPK2 and SUMO-1 to nuclear speckles (dots). CV-1 cells overexpressing Myc-HIPK2 and GFP-SUMO-1 were fixed and subjected to immunofluorescence staining with a mouse anti-Myc mAb. The red signal (HIPK2) is obtained with an anti-mouse IgG rhodamine red-conjugated secondary Ab, whereas the green signal (SUMO-1) is obtained with GFP fluorescence. Superimposing two colors (MERGE) results in a yellow signal, indicating colocalization of the two proteins.
    Figure Legend Snippet: Localization of SUMO-1-modified HIPK2 to nuclear speckles (dots). ( A ) Fractionation of SUMO-1-modified HIPK2. Cellular extracts from CV-1 cells cotransfected with GFP-SUMO-1 and Myc-HIPK2 expression vectors were fractionated. Proteins from each fraction were separated on SDS/polyacrylamide gradient (4–12%) gels and analyzed by Western blotting using a mouse anti-Myc Ab. HIPK2 and SUMO-1-modified HIPK2 are detected in the nuclear fraction (lane 3) and in the Nonidet P-40-insoluble fractions (lanes 7 and 8). SUMO-1-modified HIPK2 and unmodified HIPK2 are marked by bracket and arrowhead, respectively. ( B ) Confocal microscopic image of the nucleus showing colocalization of HIPK2 and SUMO-1 to nuclear speckles (dots). CV-1 cells overexpressing Myc-HIPK2 and GFP-SUMO-1 were fixed and subjected to immunofluorescence staining with a mouse anti-Myc mAb. The red signal (HIPK2) is obtained with an anti-mouse IgG rhodamine red-conjugated secondary Ab, whereas the green signal (SUMO-1) is obtained with GFP fluorescence. Superimposing two colors (MERGE) results in a yellow signal, indicating colocalization of the two proteins.

    Techniques Used: Modification, Fractionation, Expressing, Western Blot, Immunofluorescence, Staining, Fluorescence

    Related Articles

    Purification:

    Article Title: PTENα regulates mitophagy and maintains mitochondrial quality control
    Article Snippet: Potential interacting proteins in specific bands were evaluated with mass spectroscopy analysis. .. Purified PTENα-His (1 µg) was incubated with 3 μg GST-PRKN in PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 2 mM KH2 PO4 ) containing 0.1% NP40 supplemented with PMSF (BioDee, DE-0754), and PRKN mouse monoclonal antibody or mouse lgG (Santa Cruz Biotechnology, sc-2025) was added and incubated at 4°C for 4 h, then washed 3 times with PBS (with 0.1% NP40) followed by western blot analysis. ..

    Incubation:

    Article Title: PTENα regulates mitophagy and maintains mitochondrial quality control
    Article Snippet: Potential interacting proteins in specific bands were evaluated with mass spectroscopy analysis. .. Purified PTENα-His (1 µg) was incubated with 3 μg GST-PRKN in PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 2 mM KH2 PO4 ) containing 0.1% NP40 supplemented with PMSF (BioDee, DE-0754), and PRKN mouse monoclonal antibody or mouse lgG (Santa Cruz Biotechnology, sc-2025) was added and incubated at 4°C for 4 h, then washed 3 times with PBS (with 0.1% NP40) followed by western blot analysis. ..

    Article Title: A PGC-1α-O-GlcNAc Transferase Complex Regulates FoxO Transcription Factor Activity in Response to Glucose
    Article Snippet: Gel filtration chromatography was performed using the SMART system with a Superose 12 column (Amersham Biosciences). .. Rat liver was lysed in Tris-buffered saline with 1% Nonidet P-40 and incubated for 1 h on ice with either normal rabit IgG (Santa Cruz Biotechnology) or anti-Ogt (AL28) ( ). .. OGT Assays —The plasmid expressing recombinant OGT was a kind gift of Suzanne Walker ( ).

    Article Title: Covalent modification of the homeodomain-interacting protein kinase 2 (HIPK2) by the ubiquitin-like protein SUMO-1
    Article Snippet: The signals were detected with chemiluminescence (Supersignal; Pierce). .. For immunoprecipitation, cell extracts (500 μg) were diluted (1:10) in 1× NETN [50 mM Tris⋅HCl (pH 7.5)/5 mM EDTA/300 mM NaCl/1 mM DTT/1% Nonidet P-40] containing protease inhibitors and incubated with an anti-GFP rabbit polyclonal Ab (10 μg/ml) or anti-SUMO-1 goat Ab (Santa Cruz Biotechnology) for 1 hr at 4°C. .. After adding protein A/G plus agarose beads (Santa Cruz Biotechnology), the mixtures were further incubated for 4 hr at 4°C with mild agitation.

    Article Title: Phosphorylation of RUNX1 by Cyclin-dependent Kinase Reduces Direct Interaction with HDAC1 and HDAC3 *
    Article Snippet: After clarification at 14,000 × g for 15 min, aliquots of cell extracts were saved as “input,” and supernatants were precleared used 50 μl of 50% protein A-Sepharose. .. 200 μg of 293T or 400 μg of Jurkat or M1 total protein was then added to 1 ml of IP buffer (180 m m KCl, 0.05% Nonidet P-40, 1 m m DTT, 1 m m PMSF) and incubated with 8 μg of rabbit IgG, rabbit anti-Myc A-14 antiserum (Santa Cruz Biotechnology) for 3 h or with rabbit anti-RUNX1 antiserum (Active Motif), mouse anti-HDAC1 clone 2E10, or HDAC3 clone 3G6 (Millipore) overnight at 4 °C, followed by addition of 50 μl of protein A-Sepharose. .. The beads were then washed three times with IP buffer, and the samples were eluted in Laemmli sample buffer at 95 °C and subjected to Western blotting using mouse 9E10 anti-Myc (Santa Cruz Biotechnology), mouse M2 anti-FLAG (Sigma), rabbit anti-HDAC1 ab7028 (Abcam), mouse anti-HDAC3 clone 3G6, or rabbit anti-RUNX1 antibodies as described previously ( ).

    Article Title: Herpes Simplex Virus 2 (HSV-2) Infected Cell Proteins Are among the Most Dominant Antigens of a Live-Attenuated HSV-2 Vaccine
    Article Snippet: Background fluorescence was defined as the average ΔMFI observed in 3-population test cells incubated with naïve mouse sera. .. Immunoprecipitation-mass spectrometry analysis Two 100-mm dishes containing 107 Vero cells were mock-inoculated and two dishes were inoculated with 5 pfu per cell of HSV-2 MS. At 12 hours p.i., Vero cells were washed with ice-cold phosphate-buffered saline and lysed in 0.5 ml of an IP buffer (50 mM Tris (pH 7.4), 150 mM sodium chloride, 2 mM EDTA, 1% NP40, and 1× Halt protease inhibitor cocktail) for 2 hours at 4°C on a rotisserie, cell debris was removed by centrifugation, and supernatants were pre-cleared by incubation with Protein A/G agarose beads (SantaCruz Biotechnology) for 30 minute at 4°C. .. In the first immunoprecipitation experiment (IP Exp 1), the pre-cleared supernatants were incubated with 1.

    Western Blot:

    Article Title: PTENα regulates mitophagy and maintains mitochondrial quality control
    Article Snippet: Potential interacting proteins in specific bands were evaluated with mass spectroscopy analysis. .. Purified PTENα-His (1 µg) was incubated with 3 μg GST-PRKN in PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2 HPO4 , 2 mM KH2 PO4 ) containing 0.1% NP40 supplemented with PMSF (BioDee, DE-0754), and PRKN mouse monoclonal antibody or mouse lgG (Santa Cruz Biotechnology, sc-2025) was added and incubated at 4°C for 4 h, then washed 3 times with PBS (with 0.1% NP40) followed by western blot analysis. ..

    Article Title: Delamination of neural crest cells requires transient and reversible Wnt inhibition mediated by Dact1/2
    Article Snippet: Samples were obtained 48 hpe, lysed in NP40 buffer (150 mM NaCl, 1% NP-40, 50 mM Tris-HCl, pH 7.5 and 1 mM PMSF) and centrifuged at 20,000 g for 30 min at 4°C. .. The resulting supernatant and pellet, considered as the NP40-soluble and -insoluble fractions, respectively, were resolved by SDS-PAGE and analysed by western blot probed with rabbit polyclonal anti-Flag (R1, produced in the lab, 1:2000), mouse monoclonal anti-human SUMO1 (Santa Cruz, sc-5308, 1:2000), mouse monoclonal anti-α-tubulin (Sigma,T6119,1:1000) or rabbit polyclonal anti-PHF8 (Abcam, 36068, 1:1000). .. The α-tubulin and PHF8 proteins were used as NP40-soluble and -insoluble reference proteins, respectively.

    Article Title: The agnoprotein of Polyomavirus JC is Released by Infected Cells: Evidence for Its Cellular Uptake by Uninfected Neighboring Cells
    Article Snippet: .. The precipitated exosomes were lysed with TNN buffer containing 1% NP40, and analyzed by Western blot for the detection of agnoprotein and exosome marker Hsp70 (Santa Cruz, W27). .. The exosome depleted growth media was subjected to immunoprecipitation for agnoprotein.

    SDS Page:

    Article Title: Delamination of neural crest cells requires transient and reversible Wnt inhibition mediated by Dact1/2
    Article Snippet: Samples were obtained 48 hpe, lysed in NP40 buffer (150 mM NaCl, 1% NP-40, 50 mM Tris-HCl, pH 7.5 and 1 mM PMSF) and centrifuged at 20,000 g for 30 min at 4°C. .. The resulting supernatant and pellet, considered as the NP40-soluble and -insoluble fractions, respectively, were resolved by SDS-PAGE and analysed by western blot probed with rabbit polyclonal anti-Flag (R1, produced in the lab, 1:2000), mouse monoclonal anti-human SUMO1 (Santa Cruz, sc-5308, 1:2000), mouse monoclonal anti-α-tubulin (Sigma,T6119,1:1000) or rabbit polyclonal anti-PHF8 (Abcam, 36068, 1:1000). .. The α-tubulin and PHF8 proteins were used as NP40-soluble and -insoluble reference proteins, respectively.

    Produced:

    Article Title: Delamination of neural crest cells requires transient and reversible Wnt inhibition mediated by Dact1/2
    Article Snippet: Samples were obtained 48 hpe, lysed in NP40 buffer (150 mM NaCl, 1% NP-40, 50 mM Tris-HCl, pH 7.5 and 1 mM PMSF) and centrifuged at 20,000 g for 30 min at 4°C. .. The resulting supernatant and pellet, considered as the NP40-soluble and -insoluble fractions, respectively, were resolved by SDS-PAGE and analysed by western blot probed with rabbit polyclonal anti-Flag (R1, produced in the lab, 1:2000), mouse monoclonal anti-human SUMO1 (Santa Cruz, sc-5308, 1:2000), mouse monoclonal anti-α-tubulin (Sigma,T6119,1:1000) or rabbit polyclonal anti-PHF8 (Abcam, 36068, 1:1000). .. The α-tubulin and PHF8 proteins were used as NP40-soluble and -insoluble reference proteins, respectively.

    Marker:

    Article Title: The agnoprotein of Polyomavirus JC is Released by Infected Cells: Evidence for Its Cellular Uptake by Uninfected Neighboring Cells
    Article Snippet: .. The precipitated exosomes were lysed with TNN buffer containing 1% NP40, and analyzed by Western blot for the detection of agnoprotein and exosome marker Hsp70 (Santa Cruz, W27). .. The exosome depleted growth media was subjected to immunoprecipitation for agnoprotein.

    Immunoprecipitation:

    Article Title: Covalent modification of the homeodomain-interacting protein kinase 2 (HIPK2) by the ubiquitin-like protein SUMO-1
    Article Snippet: The signals were detected with chemiluminescence (Supersignal; Pierce). .. For immunoprecipitation, cell extracts (500 μg) were diluted (1:10) in 1× NETN [50 mM Tris⋅HCl (pH 7.5)/5 mM EDTA/300 mM NaCl/1 mM DTT/1% Nonidet P-40] containing protease inhibitors and incubated with an anti-GFP rabbit polyclonal Ab (10 μg/ml) or anti-SUMO-1 goat Ab (Santa Cruz Biotechnology) for 1 hr at 4°C. .. After adding protein A/G plus agarose beads (Santa Cruz Biotechnology), the mixtures were further incubated for 4 hr at 4°C with mild agitation.

    Article Title: Herpes Simplex Virus 2 (HSV-2) Infected Cell Proteins Are among the Most Dominant Antigens of a Live-Attenuated HSV-2 Vaccine
    Article Snippet: Background fluorescence was defined as the average ΔMFI observed in 3-population test cells incubated with naïve mouse sera. .. Immunoprecipitation-mass spectrometry analysis Two 100-mm dishes containing 107 Vero cells were mock-inoculated and two dishes were inoculated with 5 pfu per cell of HSV-2 MS. At 12 hours p.i., Vero cells were washed with ice-cold phosphate-buffered saline and lysed in 0.5 ml of an IP buffer (50 mM Tris (pH 7.4), 150 mM sodium chloride, 2 mM EDTA, 1% NP40, and 1× Halt protease inhibitor cocktail) for 2 hours at 4°C on a rotisserie, cell debris was removed by centrifugation, and supernatants were pre-cleared by incubation with Protein A/G agarose beads (SantaCruz Biotechnology) for 30 minute at 4°C. .. In the first immunoprecipitation experiment (IP Exp 1), the pre-cleared supernatants were incubated with 1.

    Mass Spectrometry:

    Article Title: Herpes Simplex Virus 2 (HSV-2) Infected Cell Proteins Are among the Most Dominant Antigens of a Live-Attenuated HSV-2 Vaccine
    Article Snippet: Background fluorescence was defined as the average ΔMFI observed in 3-population test cells incubated with naïve mouse sera. .. Immunoprecipitation-mass spectrometry analysis Two 100-mm dishes containing 107 Vero cells were mock-inoculated and two dishes were inoculated with 5 pfu per cell of HSV-2 MS. At 12 hours p.i., Vero cells were washed with ice-cold phosphate-buffered saline and lysed in 0.5 ml of an IP buffer (50 mM Tris (pH 7.4), 150 mM sodium chloride, 2 mM EDTA, 1% NP40, and 1× Halt protease inhibitor cocktail) for 2 hours at 4°C on a rotisserie, cell debris was removed by centrifugation, and supernatants were pre-cleared by incubation with Protein A/G agarose beads (SantaCruz Biotechnology) for 30 minute at 4°C. .. In the first immunoprecipitation experiment (IP Exp 1), the pre-cleared supernatants were incubated with 1.

    Protease Inhibitor:

    Article Title: Herpes Simplex Virus 2 (HSV-2) Infected Cell Proteins Are among the Most Dominant Antigens of a Live-Attenuated HSV-2 Vaccine
    Article Snippet: Background fluorescence was defined as the average ΔMFI observed in 3-population test cells incubated with naïve mouse sera. .. Immunoprecipitation-mass spectrometry analysis Two 100-mm dishes containing 107 Vero cells were mock-inoculated and two dishes were inoculated with 5 pfu per cell of HSV-2 MS. At 12 hours p.i., Vero cells were washed with ice-cold phosphate-buffered saline and lysed in 0.5 ml of an IP buffer (50 mM Tris (pH 7.4), 150 mM sodium chloride, 2 mM EDTA, 1% NP40, and 1× Halt protease inhibitor cocktail) for 2 hours at 4°C on a rotisserie, cell debris was removed by centrifugation, and supernatants were pre-cleared by incubation with Protein A/G agarose beads (SantaCruz Biotechnology) for 30 minute at 4°C. .. In the first immunoprecipitation experiment (IP Exp 1), the pre-cleared supernatants were incubated with 1.

    Article Title: An increased population of regulatory T cells improves the pathophysiology of placental ischemia in a rat model of preeclampsia
    Article Snippet: .. Placentas and cortices were removed and homogenized in RIPA buffer (PBS, 1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% SDS, and a protease inhibitor cocktail; Santa Cruz, Santa Cruz, CA), as described previously ( , ). .. The supernatant was incubated with lucigenin at a final concentration of 5 μmol/l.

    Centrifugation:

    Article Title: Herpes Simplex Virus 2 (HSV-2) Infected Cell Proteins Are among the Most Dominant Antigens of a Live-Attenuated HSV-2 Vaccine
    Article Snippet: Background fluorescence was defined as the average ΔMFI observed in 3-population test cells incubated with naïve mouse sera. .. Immunoprecipitation-mass spectrometry analysis Two 100-mm dishes containing 107 Vero cells were mock-inoculated and two dishes were inoculated with 5 pfu per cell of HSV-2 MS. At 12 hours p.i., Vero cells were washed with ice-cold phosphate-buffered saline and lysed in 0.5 ml of an IP buffer (50 mM Tris (pH 7.4), 150 mM sodium chloride, 2 mM EDTA, 1% NP40, and 1× Halt protease inhibitor cocktail) for 2 hours at 4°C on a rotisserie, cell debris was removed by centrifugation, and supernatants were pre-cleared by incubation with Protein A/G agarose beads (SantaCruz Biotechnology) for 30 minute at 4°C. .. In the first immunoprecipitation experiment (IP Exp 1), the pre-cleared supernatants were incubated with 1.

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    Santa Cruz Biotechnology nonidet p 40
    PGC-1α interacts with OGT. A , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and OGT co-immunoprecipitation ( IP ). Immunoprecipitates were  subjected to SDS-PAGE and immunoblotted ( IB ) for the presence of  PGC-1α. Membranes were then stripped and blotted for OGT. Immunoblots  are representative of three experiments.  B , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and FLAG co-immunoprecipitation. Immunoprecipitates were subjected to SDS-PAGE  and immunoblotted for the presence of OGT. Membranes were then stripped and  blotted for PGC-1α.  C , gel filtration chromatography (SMART  system, Superdex 200 column, phosphate-buffered saline, 1% Nonidet P-40  buffer) of lysates from rat liver incubated on ice for 30 min with either  normal IgG or anti-OGT antibodies (AL28). Fractions were subjected to SDS-PAGE  and blotted using anti-PGC-1α, anti-OGT (DM-17), and anti CPB. Data are  representative of two experiments.
    Nonidet P 40, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nonidet p 40/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nonidet p 40 - by Bioz Stars, 2021-05
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    97
    Santa Cruz Biotechnology ebc buffer
    Direct interaction between 4ICD and STAT5A is mediated by HER4 Y984. HEK 293T cells were transfected with the indicated expression vectors and cell lysates were prepared at 48 h. <t>post-transfection</t> in <t>EBC</t> buffer. For immunoprecipitations, 500 μg of cleared cell lysates were incubated with HER4 or STAT5A specific antibodies at 4 °C overnight. Immune complexes were recovered by adding Protein A sepharose to each immunoprecipitation reaction and incubating for 3 h at 4 °C, and finally eluted into 60 μl of NuPAGE LDS Sample Buffer with Reducing Agent. Twenty μl of eluted immunoprecipitation reactions or 20 μg of EBC lysate (Input) was probed by western blot using the indicated immunoblot (IB) antibodies.
    Ebc Buffer, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ebc buffer - by Bioz Stars, 2021-05
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    Image Search Results


    PGC-1α interacts with OGT. A , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and OGT co-immunoprecipitation ( IP ). Immunoprecipitates were  subjected to SDS-PAGE and immunoblotted ( IB ) for the presence of  PGC-1α. Membranes were then stripped and blotted for OGT. Immunoblots  are representative of three experiments.  B , Fao cells were infected  with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting  and FLAG co-immunoprecipitation. Immunoprecipitates were subjected to SDS-PAGE  and immunoblotted for the presence of OGT. Membranes were then stripped and  blotted for PGC-1α.  C , gel filtration chromatography (SMART  system, Superdex 200 column, phosphate-buffered saline, 1% Nonidet P-40  buffer) of lysates from rat liver incubated on ice for 30 min with either  normal IgG or anti-OGT antibodies (AL28). Fractions were subjected to SDS-PAGE  and blotted using anti-PGC-1α, anti-OGT (DM-17), and anti CPB. Data are  representative of two experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: A PGC-1α-O-GlcNAc Transferase Complex Regulates FoxO Transcription Factor Activity in Response to Glucose

    doi: 10.1074/jbc.M808890200

    Figure Lengend Snippet: PGC-1α interacts with OGT. A , Fao cells were infected with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting and OGT co-immunoprecipitation ( IP ). Immunoprecipitates were subjected to SDS-PAGE and immunoblotted ( IB ) for the presence of PGC-1α. Membranes were then stripped and blotted for OGT. Immunoblots are representative of three experiments. B , Fao cells were infected with Ad-FLAG-PGC-1α and treated with insulin for 1 h prior to harvesting and FLAG co-immunoprecipitation. Immunoprecipitates were subjected to SDS-PAGE and immunoblotted for the presence of OGT. Membranes were then stripped and blotted for PGC-1α. C , gel filtration chromatography (SMART system, Superdex 200 column, phosphate-buffered saline, 1% Nonidet P-40 buffer) of lysates from rat liver incubated on ice for 30 min with either normal IgG or anti-OGT antibodies (AL28). Fractions were subjected to SDS-PAGE and blotted using anti-PGC-1α, anti-OGT (DM-17), and anti CPB. Data are representative of two experiments.

    Article Snippet: Rat liver was lysed in Tris-buffered saline with 1% Nonidet P-40 and incubated for 1 h on ice with either normal rabit IgG (Santa Cruz Biotechnology) or anti-Ogt (AL28) ( ).

    Techniques: Pyrolysis Gas Chromatography, Infection, Immunoprecipitation, SDS Page, Western Blot, Filtration, Chromatography, Incubation

    Localization of SUMO-1-modified HIPK2 to nuclear speckles (dots). ( A ) Fractionation of SUMO-1-modified HIPK2. Cellular extracts from CV-1 cells cotransfected with GFP-SUMO-1 and Myc-HIPK2 expression vectors were fractionated. Proteins from each fraction were separated on SDS/polyacrylamide gradient (4–12%) gels and analyzed by Western blotting using a mouse anti-Myc Ab. HIPK2 and SUMO-1-modified HIPK2 are detected in the nuclear fraction (lane 3) and in the Nonidet P-40-insoluble fractions (lanes 7 and 8). SUMO-1-modified HIPK2 and unmodified HIPK2 are marked by bracket and arrowhead, respectively. ( B ) Confocal microscopic image of the nucleus showing colocalization of HIPK2 and SUMO-1 to nuclear speckles (dots). CV-1 cells overexpressing Myc-HIPK2 and GFP-SUMO-1 were fixed and subjected to immunofluorescence staining with a mouse anti-Myc mAb. The red signal (HIPK2) is obtained with an anti-mouse IgG rhodamine red-conjugated secondary Ab, whereas the green signal (SUMO-1) is obtained with GFP fluorescence. Superimposing two colors (MERGE) results in a yellow signal, indicating colocalization of the two proteins.

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

    Article Title: Covalent modification of the homeodomain-interacting protein kinase 2 (HIPK2) by the ubiquitin-like protein SUMO-1

    doi:

    Figure Lengend Snippet: Localization of SUMO-1-modified HIPK2 to nuclear speckles (dots). ( A ) Fractionation of SUMO-1-modified HIPK2. Cellular extracts from CV-1 cells cotransfected with GFP-SUMO-1 and Myc-HIPK2 expression vectors were fractionated. Proteins from each fraction were separated on SDS/polyacrylamide gradient (4–12%) gels and analyzed by Western blotting using a mouse anti-Myc Ab. HIPK2 and SUMO-1-modified HIPK2 are detected in the nuclear fraction (lane 3) and in the Nonidet P-40-insoluble fractions (lanes 7 and 8). SUMO-1-modified HIPK2 and unmodified HIPK2 are marked by bracket and arrowhead, respectively. ( B ) Confocal microscopic image of the nucleus showing colocalization of HIPK2 and SUMO-1 to nuclear speckles (dots). CV-1 cells overexpressing Myc-HIPK2 and GFP-SUMO-1 were fixed and subjected to immunofluorescence staining with a mouse anti-Myc mAb. The red signal (HIPK2) is obtained with an anti-mouse IgG rhodamine red-conjugated secondary Ab, whereas the green signal (SUMO-1) is obtained with GFP fluorescence. Superimposing two colors (MERGE) results in a yellow signal, indicating colocalization of the two proteins.

    Article Snippet: For immunoprecipitation, cell extracts (500 μg) were diluted (1:10) in 1× NETN [50 mM Tris⋅HCl (pH 7.5)/5 mM EDTA/300 mM NaCl/1 mM DTT/1% Nonidet P-40] containing protease inhibitors and incubated with an anti-GFP rabbit polyclonal Ab (10 μg/ml) or anti-SUMO-1 goat Ab (Santa Cruz Biotechnology) for 1 hr at 4°C.

    Techniques: Modification, Fractionation, Expressing, Western Blot, Immunofluorescence, Staining, Fluorescence

    Direct interaction between 4ICD and STAT5A is mediated by HER4 Y984. HEK 293T cells were transfected with the indicated expression vectors and cell lysates were prepared at 48 h. post-transfection in EBC buffer. For immunoprecipitations, 500 μg of cleared cell lysates were incubated with HER4 or STAT5A specific antibodies at 4 °C overnight. Immune complexes were recovered by adding Protein A sepharose to each immunoprecipitation reaction and incubating for 3 h at 4 °C, and finally eluted into 60 μl of NuPAGE LDS Sample Buffer with Reducing Agent. Twenty μl of eluted immunoprecipitation reactions or 20 μg of EBC lysate (Input) was probed by western blot using the indicated immunoblot (IB) antibodies.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Direct coupling of the HER4 intracellular domain (4ICD) and STAT5A signaling is required to induce mammary epithelial cell differentiation

    doi: 10.1016/j.bbrep.2016.07.015

    Figure Lengend Snippet: Direct interaction between 4ICD and STAT5A is mediated by HER4 Y984. HEK 293T cells were transfected with the indicated expression vectors and cell lysates were prepared at 48 h. post-transfection in EBC buffer. For immunoprecipitations, 500 μg of cleared cell lysates were incubated with HER4 or STAT5A specific antibodies at 4 °C overnight. Immune complexes were recovered by adding Protein A sepharose to each immunoprecipitation reaction and incubating for 3 h at 4 °C, and finally eluted into 60 μl of NuPAGE LDS Sample Buffer with Reducing Agent. Twenty μl of eluted immunoprecipitation reactions or 20 μg of EBC lysate (Input) was probed by western blot using the indicated immunoblot (IB) antibodies.

    Article Snippet: At 2 days post-transfection, cell lysates were prepared in EBC buffer (50 mM Tris, pH 7.5, 50 mM Tris, pH 7.5, 0.5% NP-40, with 1 mM phenylmethylsulfonyl fluoride) and 500 μg of lysate was immunoprecipitated overnight at 4 °C with antibodies directed against HER4 (Santa Cruz; sc-283) or STAT5 (Santa Cruz; sc-835).

    Techniques: Transfection, Expressing, Incubation, Immunoprecipitation, Western Blot