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Roche nonidet lysis buffer
Nonidet Lysis 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/nonidet lysis buffer/product/Roche
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
nonidet lysis buffer - by Bioz Stars, 2020-04
86/100 stars

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Transfection:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: Thirty-six hours after transfection, cells were starved in 1 ml of methionine and cysteine-free DMEM for 1 h, pulsed for 30 min with 1.3 MBq/well Easy TagTM Expre35S Protein Labeling Mix (PerkinElmer) containing 35 S-labeled methionine and cysteine, and then harvested or washed with cold PBS and cultured in 1 ml of chase medium (DMEM containing 200 m m methionine and 200 m m cysteine). .. After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min.

Pulse Chase:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: For the pulse-chase of 3 h, the chase medium was supplemented with 10% (v/v) fetal bovine serum. .. After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min.

Protease Inhibitor:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: .. After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min. .. The supernatants from culture medium and cell lysate samples were precleared with rabbit IgG bound to 50 μl of 50% (v/v) protein A-Sepharose for at least 1 h at 4 °C, and then neuroserpin proteins were immunoprecipitated in parallel either with a purified antibody for total neuroserpin or with a polymer-specific antibody overnight at 4 °C.

Co-Immunoprecipitation Assay:

Article Title: The translation elongation factor eEF1A1 couples transcription to translation during heat shock response
Article Snippet: .. Co-immunoprecipitation (co-IP) Cells were lysed in nonidet lysis buffer (25 mM Hepes, pH 7.9, 100 mM NaCl, 5 mM EDTA, 0.5% nonidet P40, 1× complete [Roche, Bransburg, NJ, USA], and phosphatase inhibitors [Roche, Bransburg, NJ, USA]) for 1 hr at 4°C. .. Lysates were clarified of non-protein components by centrifugation for 5 min at 14,000×g and pre-cleared with 25 μl of Dynabeads protein G (Life Technologies, Van Allen Way Carlsbad, CA, USA) washed three times with B150 buffer (20 mM Tris, pH7.4, 1 mM EDTA, 10% glycerol, 150 mM NaCl and 0.1% Triton) for 1 hr at 4°C.

Labeling:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: Paragraph title: Metabolic Labeling and Immunoprecipitation ... After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min.

Purification:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min. .. The supernatants from culture medium and cell lysate samples were precleared with rabbit IgG bound to 50 μl of 50% (v/v) protein A-Sepharose for at least 1 h at 4 °C, and then neuroserpin proteins were immunoprecipitated in parallel either with a purified antibody for total neuroserpin or with a polymer-specific antibody overnight at 4 °C.

Immunoprecipitation:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: Paragraph title: Metabolic Labeling and Immunoprecipitation ... After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min.

Incubation:

Article Title: The translation elongation factor eEF1A1 couples transcription to translation during heat shock response
Article Snippet: Co-immunoprecipitation (co-IP) Cells were lysed in nonidet lysis buffer (25 mM Hepes, pH 7.9, 100 mM NaCl, 5 mM EDTA, 0.5% nonidet P40, 1× complete [Roche, Bransburg, NJ, USA], and phosphatase inhibitors [Roche, Bransburg, NJ, USA]) for 1 hr at 4°C. .. 1 mg of pre-cleared protein was IP overnight at 4°C with 5 μg of anti-eEF1A1 (Millipore, Billerica, MA, USA) or normal rabbit or mouse IgG (Abcam, Cambridge, MA, USA) followed by a 1-hr incubation with 50 μl of Dynabeads protein G (Life Technologies, Van Allen Way Carlsbad, CA, USA).

Cell Culture:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: Thirty-six hours after transfection, cells were starved in 1 ml of methionine and cysteine-free DMEM for 1 h, pulsed for 30 min with 1.3 MBq/well Easy TagTM Expre35S Protein Labeling Mix (PerkinElmer) containing 35 S-labeled methionine and cysteine, and then harvested or washed with cold PBS and cultured in 1 ml of chase medium (DMEM containing 200 m m methionine and 200 m m cysteine). .. After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min.

Lysis:

Article Title: Endoplasmic Reticulum-associated Degradation (ERAD) and Autophagy Cooperate to Degrade Polymerogenic Mutant Serpins *
Article Snippet: .. After the chase period, the culture medium was collected and centrifuged at 2000 rpm and 4 °C for 11 min, and the cells were harvested by adding 0.5 ml/well Nonidet lysis buffer (150 m m NaCl, 50 m m Tris-Cl, pH 7.5, 1% v/v Nonidet P-40) containing a protease inhibitor mixture (Complete; EDTA-free protease inhibitor mixture tablets (Roche Applied Science)), scraping and vortexing for 3 × 3 s, and then centrifuging at 16,000 rpm and 4 °C for 16 min. .. The supernatants from culture medium and cell lysate samples were precleared with rabbit IgG bound to 50 μl of 50% (v/v) protein A-Sepharose for at least 1 h at 4 °C, and then neuroserpin proteins were immunoprecipitated in parallel either with a purified antibody for total neuroserpin or with a polymer-specific antibody overnight at 4 °C.

Article Title: The translation elongation factor eEF1A1 couples transcription to translation during heat shock response
Article Snippet: .. Co-immunoprecipitation (co-IP) Cells were lysed in nonidet lysis buffer (25 mM Hepes, pH 7.9, 100 mM NaCl, 5 mM EDTA, 0.5% nonidet P40, 1× complete [Roche, Bransburg, NJ, USA], and phosphatase inhibitors [Roche, Bransburg, NJ, USA]) for 1 hr at 4°C. .. Lysates were clarified of non-protein components by centrifugation for 5 min at 14,000×g and pre-cleared with 25 μl of Dynabeads protein G (Life Technologies, Van Allen Way Carlsbad, CA, USA) washed three times with B150 buffer (20 mM Tris, pH7.4, 1 mM EDTA, 10% glycerol, 150 mM NaCl and 0.1% Triton) for 1 hr at 4°C.

Centrifugation:

Article Title: The translation elongation factor eEF1A1 couples transcription to translation during heat shock response
Article Snippet: Co-immunoprecipitation (co-IP) Cells were lysed in nonidet lysis buffer (25 mM Hepes, pH 7.9, 100 mM NaCl, 5 mM EDTA, 0.5% nonidet P40, 1× complete [Roche, Bransburg, NJ, USA], and phosphatase inhibitors [Roche, Bransburg, NJ, USA]) for 1 hr at 4°C. .. Lysates were clarified of non-protein components by centrifugation for 5 min at 14,000×g and pre-cleared with 25 μl of Dynabeads protein G (Life Technologies, Van Allen Way Carlsbad, CA, USA) washed three times with B150 buffer (20 mM Tris, pH7.4, 1 mM EDTA, 10% glycerol, 150 mM NaCl and 0.1% Triton) for 1 hr at 4°C.

SDS Page:

Article Title: The translation elongation factor eEF1A1 couples transcription to translation during heat shock response
Article Snippet: Co-immunoprecipitation (co-IP) Cells were lysed in nonidet lysis buffer (25 mM Hepes, pH 7.9, 100 mM NaCl, 5 mM EDTA, 0.5% nonidet P40, 1× complete [Roche, Bransburg, NJ, USA], and phosphatase inhibitors [Roche, Bransburg, NJ, USA]) for 1 hr at 4°C. .. After five washes in B150, the proteins were resolved by SDS-PAGE and analyzed by immunoblotting.

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  • 94
    Roche nondenaturing np 40 lysis buffer
    Western blot analysis of pyrin S205 dephosphorylation in BMDMs. LPS-primed BMDMs were left uninfected, infected with Y. pseudotuberculosis strain IP6ΔM harboring an empty vector or IP6ΔM/pYopE or intoxicated with TcdB for 90 min. (A and B) Lysates were processed with <t>nondenaturing</t> <t>NP-40</t> lysis buffer and separated into soluble and insoluble fractions by centrifugation. (C) Lysates were processed with denaturing RIPA lysis buffer, and the soluble fractions were collected. Samples were subjected to Western blotting using antibodies to pSer205 of pyrin, total pyrin, or β-actin.
    Nondenaturing Np 40 Lysis Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nondenaturing np 40 lysis buffer/product/Roche
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nondenaturing np 40 lysis buffer - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    99
    Roche np 40 lysis buffer
    CsCl density gradient analysis of hepatitis B viral particles. (A and B) CsCl density gradient analysis of viral particles in patient sera. One hundred-microliter volumes of serum mixture from patients 37, 38, 14, and 35 (25 μl each) and 100 μl serum from patient 17 were separated by CsCl density gradient centrifugation (2 ml of 1.18 g/cm 3 CsCl solution in the upper layer and 2.9 ml of 1.33 g/cm 3 CsCl solution in the lower layer). Viral DNA in each fraction was extracted and detected by Southern blotting. (C to G) CsCl density gradient analysis of viral particles treated with detergent or anti-HBcAg antibody (Ab). Concentrated HepAD38 cell culture supernatant (250 μl each) (via ultrafiltration) was either mixed with anti-HBcAg antibody (10 μl) followed by incubation without (C) or with <t>NP-40</t> (final concentration, 1%) (D) for 1 h at room temperature and 4 h on ice or treated with only NP-40 (G) and then fractionated by CsCl density gradient ultracentrifugation. Sera from CHB patient 46 either left untreated (E) or treated with NP-40 (final concentration, 1%) (F) were fractionated by CsCl density gradient ultracentrifugation. Viral DNA in each fraction was extracted and subjected to Southern blot analyses.
    Np 40 Lysis Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 137 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/np 40 lysis buffer/product/Roche
    Average 99 stars, based on 137 article reviews
    Price from $9.99 to $1999.99
    np 40 lysis buffer - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    93
    Roche immunoprecipitation ip buffer
    Mode of inhibition of TNFR1 signaling by zafirlukast and triclabendazole (A) Effect of zafirlukast and triclabendazole (200 μM) on TNFR1-LTα interaction was determined by <t>co-immunoprecipitation</t> with anti-FLAG–conjugated agarose beads. (B) Oligomeric states of PLAD were determined by analytical size-exclusion chromatography. Dimer peak is indicated. Open triangle indicates non-specific protein. (C) Effect of zafirlukast and triclabendazole on PLAD-PLAD interactions was determined by Native-PAGE. Open circle indicates nonspecific band. (D) Docking scores for zafirlukast and triclabendazole on TNFR1 chain B (PDB: 1NCF) interfacial residues. Boltzmann-weighted scores were calculated by multiplying the probability of contacting a given residue by the mean Boltzmann-weighted predicted free energy for each pose contacting that residue.
    Immunoprecipitation Ip Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation ip buffer/product/Roche
    Average 93 stars, based on 42 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitation ip buffer - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    Image Search Results


    Western blot analysis of pyrin S205 dephosphorylation in BMDMs. LPS-primed BMDMs were left uninfected, infected with Y. pseudotuberculosis strain IP6ΔM harboring an empty vector or IP6ΔM/pYopE or intoxicated with TcdB for 90 min. (A and B) Lysates were processed with nondenaturing NP-40 lysis buffer and separated into soluble and insoluble fractions by centrifugation. (C) Lysates were processed with denaturing RIPA lysis buffer, and the soluble fractions were collected. Samples were subjected to Western blotting using antibodies to pSer205 of pyrin, total pyrin, or β-actin.

    Journal: Infection and Immunity

    Article Title: Characterization of Pyrin Dephosphorylation and Inflammasome Activation in Macrophages as Triggered by the Yersinia Effectors YopE and YopT

    doi: 10.1128/IAI.00822-18

    Figure Lengend Snippet: Western blot analysis of pyrin S205 dephosphorylation in BMDMs. LPS-primed BMDMs were left uninfected, infected with Y. pseudotuberculosis strain IP6ΔM harboring an empty vector or IP6ΔM/pYopE or intoxicated with TcdB for 90 min. (A and B) Lysates were processed with nondenaturing NP-40 lysis buffer and separated into soluble and insoluble fractions by centrifugation. (C) Lysates were processed with denaturing RIPA lysis buffer, and the soluble fractions were collected. Samples were subjected to Western blotting using antibodies to pSer205 of pyrin, total pyrin, or β-actin.

    Article Snippet: To obtain cell lysates, BMDMs were lysed using nondenaturing NP-40 lysis buffer (1% NP-40, 150 mM NaCl, and 50 mM Tris-HCl, pH 8.0) or denaturing RIPA buffer (50 mM Tris, pH 8.0, 150 mM NaCl, 1% NP-40, 0.5% deoxycholic acid [DOC], and 0.1% SDS) with protease inhibitor cocktail (Roche).

    Techniques: Western Blot, De-Phosphorylation Assay, Infection, Plasmid Preparation, Lysis, Centrifugation

    CsCl density gradient analysis of hepatitis B viral particles. (A and B) CsCl density gradient analysis of viral particles in patient sera. One hundred-microliter volumes of serum mixture from patients 37, 38, 14, and 35 (25 μl each) and 100 μl serum from patient 17 were separated by CsCl density gradient centrifugation (2 ml of 1.18 g/cm 3 CsCl solution in the upper layer and 2.9 ml of 1.33 g/cm 3 CsCl solution in the lower layer). Viral DNA in each fraction was extracted and detected by Southern blotting. (C to G) CsCl density gradient analysis of viral particles treated with detergent or anti-HBcAg antibody (Ab). Concentrated HepAD38 cell culture supernatant (250 μl each) (via ultrafiltration) was either mixed with anti-HBcAg antibody (10 μl) followed by incubation without (C) or with NP-40 (final concentration, 1%) (D) for 1 h at room temperature and 4 h on ice or treated with only NP-40 (G) and then fractionated by CsCl density gradient ultracentrifugation. Sera from CHB patient 46 either left untreated (E) or treated with NP-40 (final concentration, 1%) (F) were fractionated by CsCl density gradient ultracentrifugation. Viral DNA in each fraction was extracted and subjected to Southern blot analyses.

    Journal: Journal of Virology

    Article Title: Extracellular Hepatitis B Virus RNAs Are Heterogeneous in Length and Circulate as Capsid-Antibody Complexes in Addition to Virions in Chronic Hepatitis B Patients

    doi: 10.1128/JVI.00798-18

    Figure Lengend Snippet: CsCl density gradient analysis of hepatitis B viral particles. (A and B) CsCl density gradient analysis of viral particles in patient sera. One hundred-microliter volumes of serum mixture from patients 37, 38, 14, and 35 (25 μl each) and 100 μl serum from patient 17 were separated by CsCl density gradient centrifugation (2 ml of 1.18 g/cm 3 CsCl solution in the upper layer and 2.9 ml of 1.33 g/cm 3 CsCl solution in the lower layer). Viral DNA in each fraction was extracted and detected by Southern blotting. (C to G) CsCl density gradient analysis of viral particles treated with detergent or anti-HBcAg antibody (Ab). Concentrated HepAD38 cell culture supernatant (250 μl each) (via ultrafiltration) was either mixed with anti-HBcAg antibody (10 μl) followed by incubation without (C) or with NP-40 (final concentration, 1%) (D) for 1 h at room temperature and 4 h on ice or treated with only NP-40 (G) and then fractionated by CsCl density gradient ultracentrifugation. Sera from CHB patient 46 either left untreated (E) or treated with NP-40 (final concentration, 1%) (F) were fractionated by CsCl density gradient ultracentrifugation. Viral DNA in each fraction was extracted and subjected to Southern blot analyses.

    Article Snippet: To isolate intracellular capsid-associated viral RNA, HepAD38 cells were lysed in NP-40 lysis buffer (50 mM Tris-Cl [pH 7.8], 1 mM EDTA, 1% NP-40), and cytoplasmic lysates were incubated with CaCl2 (final concentration, 5 mM) and micrococcal nuclease (MNase) (Roche) (final concentration, 15 U/ml) at 37°C for 1 h to remove nucleic acids outside nucleocapsids.

    Techniques: Gradient Centrifugation, Southern Blot, Cell Culture, Incubation, Concentration Assay

    Isolation of HBV NCs from MNase-treated lysates by linear sucrose gradient centrifugation and detection of NC-associated viral DNA, RNA, and capsid protein. Induced HepAD38 cells were lysed by using NP-40, and the cytoplasmic lysate was treated with MNase.

    Journal: Journal of Virology

    Article Title: Maturation-Associated Destabilization of Hepatitis B Virus Nucleocapsid

    doi: 10.1128/JVI.01912-13

    Figure Lengend Snippet: Isolation of HBV NCs from MNase-treated lysates by linear sucrose gradient centrifugation and detection of NC-associated viral DNA, RNA, and capsid protein. Induced HepAD38 cells were lysed by using NP-40, and the cytoplasmic lysate was treated with MNase.

    Article Snippet: HepAD38 cells induced for 20 days without Tet were lysed in NP-40 lysis buffer (50 mM Tris-HCl [pH 8.0], 1 mM EDTA, 1% NP-40) containing a protease inhibitor cocktail (Roche) and briefly centrifuged at 14,000 rpm to remove the nuclei and cell debris.

    Techniques: Isolation, Gradient Centrifugation

    Mode of inhibition of TNFR1 signaling by zafirlukast and triclabendazole (A) Effect of zafirlukast and triclabendazole (200 μM) on TNFR1-LTα interaction was determined by co-immunoprecipitation with anti-FLAG–conjugated agarose beads. (B) Oligomeric states of PLAD were determined by analytical size-exclusion chromatography. Dimer peak is indicated. Open triangle indicates non-specific protein. (C) Effect of zafirlukast and triclabendazole on PLAD-PLAD interactions was determined by Native-PAGE. Open circle indicates nonspecific band. (D) Docking scores for zafirlukast and triclabendazole on TNFR1 chain B (PDB: 1NCF) interfacial residues. Boltzmann-weighted scores were calculated by multiplying the probability of contacting a given residue by the mean Boltzmann-weighted predicted free energy for each pose contacting that residue.

    Journal: SLAS discovery : advancing life sciences R & D

    Article Title: An innovative high-throughput screening approach for discovery of small molecules that inhibit TNF Receptors

    doi: 10.1177/2472555217706478

    Figure Lengend Snippet: Mode of inhibition of TNFR1 signaling by zafirlukast and triclabendazole (A) Effect of zafirlukast and triclabendazole (200 μM) on TNFR1-LTα interaction was determined by co-immunoprecipitation with anti-FLAG–conjugated agarose beads. (B) Oligomeric states of PLAD were determined by analytical size-exclusion chromatography. Dimer peak is indicated. Open triangle indicates non-specific protein. (C) Effect of zafirlukast and triclabendazole on PLAD-PLAD interactions was determined by Native-PAGE. Open circle indicates nonspecific band. (D) Docking scores for zafirlukast and triclabendazole on TNFR1 chain B (PDB: 1NCF) interfacial residues. Boltzmann-weighted scores were calculated by multiplying the probability of contacting a given residue by the mean Boltzmann-weighted predicted free energy for each pose contacting that residue.

    Article Snippet: HEK293 and transiently transfected cells were washed three times with ice-cold PBS before lysis with immunoprecipitation (IP) buffer (20 mM Tris-HCl, 1 mM EDTA, 100 mM NaCl, pH 7.5 and 0.5% NP-40) containing complete protease inhibitors cocktail (Roche).

    Techniques: Inhibition, Immunoprecipitation, Size-exclusion Chromatography, Clear Native PAGE