Review



tris-hcl buffer (ph 8.0) containing nacl  (Spectrum Labs)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 90
      Buy from Supplier

    Structured Review

    Spectrum Labs tris-hcl buffer (ph 8.0) containing nacl
    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Tris Hcl Buffer (Ph 8.0) Containing Nacl, supplied by Spectrum Labs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tris-hcl buffer (ph 8.0) containing nacl/product/Spectrum Labs
    Average 90 stars, based on 1 article reviews
    tris-hcl buffer (ph 8.0) containing nacl - by Bioz Stars, 2026-02
    90/100 stars

    Images

    1) Product Images from "Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog"

    Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

    Journal: The Journal of Biological Chemistry

    doi: 10.1016/j.jbc.2024.107763

    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Figure Legend Snippet: Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

    Techniques Used: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

    Effect of mouse chymotrypsin C (CTRC) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRC in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.
    Figure Legend Snippet: Effect of mouse chymotrypsin C (CTRC) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRC in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

    Techniques Used: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

    Effect of mouse chymotrypsin B1 (CTRB1) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRB1 in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.
    Figure Legend Snippet: Effect of mouse chymotrypsin B1 (CTRB1) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRB1 in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

    Techniques Used: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

    Effect of the T341K mutation on the degradation of mouse PNLIP variants by mouse T7 trypsin. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Figure Legend Snippet: Effect of the T341K mutation on the degradation of mouse PNLIP variants by mouse T7 trypsin. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

    Techniques Used: Mutagenesis, Purification, Concentration Assay, Incubation, SDS Page, Staining



    Similar Products

    ice-cold homogenization buffer containing 50 mm tris-hcl, 150 mm nacl, 0.1% sds, 1% np-40, and 0.5% sodium deoxycholate (ph 8.0)  (FUJIFILM)
    90
    FUJIFILM ice-cold homogenization buffer containing 50 mm tris-hcl, 150 mm nacl, 0.1% sds, 1% np-40, and 0.5% sodium deoxycholate (ph 8.0)
    Ice Cold Homogenization Buffer Containing 50 Mm Tris Hcl, 150 Mm Nacl, 0.1% Sds, 1% Np 40, And 0.5% Sodium Deoxycholate (Ph 8.0), supplied by FUJIFILM, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ice-cold homogenization buffer containing 50 mm tris-hcl, 150 mm nacl, 0.1% sds, 1% np-40, and 0.5% sodium deoxycholate (ph 8.0)/product/FUJIFILM
    Average 90 stars, based on 1 article reviews
    ice-cold homogenization buffer containing 50 mm tris-hcl, 150 mm nacl, 0.1% sds, 1% np-40, and 0.5% sodium deoxycholate (ph 8.0) - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    lysis buffer containing 25 mm tris-hcl, ph 8.0, 137 mm nacl, 2.7 mm kcl, 1% triton x-100 and protease inhibitor mixture  (Thermo Fisher)
    90
    Thermo Fisher lysis buffer containing 25 mm tris-hcl, ph 8.0, 137 mm nacl, 2.7 mm kcl, 1% triton x-100 and protease inhibitor mixture
    Lysis Buffer Containing 25 Mm Tris Hcl, Ph 8.0, 137 Mm Nacl, 2.7 Mm Kcl, 1% Triton X 100 And Protease Inhibitor Mixture, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lysis buffer containing 25 mm tris-hcl, ph 8.0, 137 mm nacl, 2.7 mm kcl, 1% triton x-100 and protease inhibitor mixture/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    lysis buffer containing 25 mm tris-hcl, ph 8.0, 137 mm nacl, 2.7 mm kcl, 1% triton x-100 and protease inhibitor mixture - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    ripa buffer containing tris hcl ph 8.0, nacl  (Merck & Co)
    90
    Merck & Co ripa buffer containing tris hcl ph 8.0, nacl
    Ripa Buffer Containing Tris Hcl Ph 8.0, Nacl, supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ripa buffer containing tris hcl ph 8.0, nacl/product/Merck & Co
    Average 90 stars, based on 1 article reviews
    ripa buffer containing tris hcl ph 8.0, nacl - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    tris-hcl buffer (ph 8.0) containing nacl, dithiothreitol, protease-inhibitor cocktail  (Nacalai)
    90
    Nacalai tris-hcl buffer (ph 8.0) containing nacl, dithiothreitol, protease-inhibitor cocktail
    Tris Hcl Buffer (Ph 8.0) Containing Nacl, Dithiothreitol, Protease Inhibitor Cocktail, supplied by Nacalai, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tris-hcl buffer (ph 8.0) containing nacl, dithiothreitol, protease-inhibitor cocktail/product/Nacalai
    Average 90 stars, based on 1 article reviews
    tris-hcl buffer (ph 8.0) containing nacl, dithiothreitol, protease-inhibitor cocktail - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    tris-hcl buffer (ph 8.0) containing nacl  (Spectrum Labs)
    90
    Spectrum Labs tris-hcl buffer (ph 8.0) containing nacl
    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Tris Hcl Buffer (Ph 8.0) Containing Nacl, supplied by Spectrum Labs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tris-hcl buffer (ph 8.0) containing nacl/product/Spectrum Labs
    Average 90 stars, based on 1 article reviews
    tris-hcl buffer (ph 8.0) containing nacl - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    ripa lysis buffer containing 50 mm tris–hcl (ph 8.0), 150 mm nacl, 1.0% nonidet p-40, 0.5% sodium deoxycholate, 0.1% sds  (Nacalai)
    90
    Nacalai ripa lysis buffer containing 50 mm tris–hcl (ph 8.0), 150 mm nacl, 1.0% nonidet p-40, 0.5% sodium deoxycholate, 0.1% sds
    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Ripa Lysis Buffer Containing 50 Mm Tris–Hcl (Ph 8.0), 150 Mm Nacl, 1.0% Nonidet P 40, 0.5% Sodium Deoxycholate, 0.1% Sds, supplied by Nacalai, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ripa lysis buffer containing 50 mm tris–hcl (ph 8.0), 150 mm nacl, 1.0% nonidet p-40, 0.5% sodium deoxycholate, 0.1% sds/product/Nacalai
    Average 90 stars, based on 1 article reviews
    ripa lysis buffer containing 50 mm tris–hcl (ph 8.0), 150 mm nacl, 1.0% nonidet p-40, 0.5% sodium deoxycholate, 0.1% sds - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    lysis buffer (50 tris-hcl, nacl, 1% sds, ph 8.0) containing protease inhibitors and phosphorylation inhibitors  (Thermo Fisher)
    90
    Thermo Fisher lysis buffer (50 tris-hcl, nacl, 1% sds, ph 8.0) containing protease inhibitors and phosphorylation inhibitors
    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Lysis Buffer (50 Tris Hcl, Nacl, 1% Sds, Ph 8.0) Containing Protease Inhibitors And Phosphorylation Inhibitors, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lysis buffer (50 tris-hcl, nacl, 1% sds, ph 8.0) containing protease inhibitors and phosphorylation inhibitors/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    lysis buffer (50 tris-hcl, nacl, 1% sds, ph 8.0) containing protease inhibitors and phosphorylation inhibitors - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    anti-bleaching buffer containing tris-hcl ph 8.0, nacl, 2×ssc, trolox  (Millipore)
    90
    Millipore anti-bleaching buffer containing tris-hcl ph 8.0, nacl, 2×ssc, trolox
    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Anti Bleaching Buffer Containing Tris Hcl Ph 8.0, Nacl, 2×Ssc, Trolox, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti-bleaching buffer containing tris-hcl ph 8.0, nacl, 2×ssc, trolox/product/Millipore
    Average 90 stars, based on 1 article reviews
    anti-bleaching buffer containing tris-hcl ph 8.0, nacl, 2×ssc, trolox - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    buffer containing tris-hcl ph 8.0, nacl n-dodecyl-β-d-maltopyranoside  (Anatrace)
    90
    Anatrace buffer containing tris-hcl ph 8.0, nacl n-dodecyl-β-d-maltopyranoside
    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Buffer Containing Tris Hcl Ph 8.0, Nacl N Dodecyl β D Maltopyranoside, supplied by Anatrace, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/buffer containing tris-hcl ph 8.0, nacl n-dodecyl-β-d-maltopyranoside/product/Anatrace
    Average 90 stars, based on 1 article reviews
    buffer containing tris-hcl ph 8.0, nacl n-dodecyl-β-d-maltopyranoside - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    radioimmunoprecipitation assay buffer containing protease inhibitors, 50 mm tris–hcl, ph 8.0, 150 mm nacl, 1% np-40  (R&D Systems)
    90
    R&D Systems radioimmunoprecipitation assay buffer containing protease inhibitors, 50 mm tris–hcl, ph 8.0, 150 mm nacl, 1% np-40
    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 <t>M</t> <t>Tris-HCl</t> (pH 8.0), 50 mM <t>NaCl,</t> and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.
    Radioimmunoprecipitation Assay Buffer Containing Protease Inhibitors, 50 Mm Tris–Hcl, Ph 8.0, 150 Mm Nacl, 1% Np 40, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/radioimmunoprecipitation assay buffer containing protease inhibitors, 50 mm tris–hcl, ph 8.0, 150 mm nacl, 1% np-40/product/R&D Systems
    Average 90 stars, based on 1 article reviews
    radioimmunoprecipitation assay buffer containing protease inhibitors, 50 mm tris–hcl, ph 8.0, 150 mm nacl, 1% np-40 - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

    Journal: The Journal of Biological Chemistry

    Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

    doi: 10.1016/j.jbc.2024.107763

    Figure Lengend Snippet: Effect of mouse cationic T7 trypsin on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , Representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

    Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

    Techniques: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

    Effect of mouse chymotrypsin C (CTRC) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRC in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

    Journal: The Journal of Biological Chemistry

    Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

    doi: 10.1016/j.jbc.2024.107763

    Figure Lengend Snippet: Effect of mouse chymotrypsin C (CTRC) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRC in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

    Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

    Techniques: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

    Effect of mouse chymotrypsin B1 (CTRB1) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRB1 in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

    Journal: The Journal of Biological Chemistry

    Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

    doi: 10.1016/j.jbc.2024.107763

    Figure Lengend Snippet: Effect of mouse chymotrypsin B1 (CTRB1) on mouse PNLIP variants. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse CTRB1 in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

    Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

    Techniques: Purification, Mutagenesis, Concentration Assay, Incubation, SDS Page, Staining

    Effect of the T341K mutation on the degradation of mouse PNLIP variants by mouse T7 trypsin. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

    Journal: The Journal of Biological Chemistry

    Article Title: Modeling protease-sensitive human pancreatic lipase mutations in the mouse ortholog

    doi: 10.1016/j.jbc.2024.107763

    Figure Lengend Snippet: Effect of the T341K mutation on the degradation of mouse PNLIP variants by mouse T7 trypsin. Purified wild-type and mutant PNLIP proteins at a final concentration of 2 μM were incubated at 37 °C with 200 nM mouse T7 trypsin in 0.1 M Tris-HCl (pH 8.0), 50 mM NaCl, and 1 mM CaCl 2 (final concentrations). At the indicated times, 75 μl aliquots were precipitated with 10% trichloroacetic acid (final concentration) and analyzed by reducing SDS-PAGE and Coomassie Blue staining. A , representative gels of three experiments are shown. B , representative evaluation of PNLIP band intensities. Mean ± SD of three experiments are shown. Statistical significance relative to the wild-type value was calculated at 60 min ∗∗ p ≤ 0.01.

    Article Snippet: Fractions containing pure PNLIP protein were pooled and dialyzed against 3 × 1000 ml of 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM NaCl using Spectra/Por Float-A-Lyzer Dialysis devices (Spectrum Labs) with molecular weight cut-off of 10 kDa.

    Techniques: Mutagenesis, Purification, Concentration Assay, Incubation, SDS Page, Staining