hepes Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Millipore hepes buffer
    (A) Structure of CN-PPV polymer and the absorption (dashed) and emission (solid) spectra of CN-PPV Pdot (black) and CN-PPV <t>Pdot-streptavidin</t> conjugate (CNPPV-Strep, red) in a 0.1% PEG 20 mM <t>HEPES</t> buffer. (B) The absorption and emission spectra of Qdot
    Hepes Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 5220 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes buffer/product/Millipore
    Average 95 stars, based on 5220 article reviews
    Price from $9.99 to $1999.99
    hepes buffer - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    90
    Thermo Fisher hepes
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Hepes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 41613 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Thermo Fisher
    Average 90 stars, based on 41613 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    77
    Amresco n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid Hepes, supplied by Amresco, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes/product/Amresco
    Average 77 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes - by Bioz Stars, 2020-02
    77/100 stars
      Buy from Supplier

    91
    Millipore n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid Hepes, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes/product/Millipore
    Average 91 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes - by Bioz Stars, 2020-02
    91/100 stars
      Buy from Supplier

    78
    Merck KGaA n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 78/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid/product/Merck KGaA
    Average 78 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid - by Bioz Stars, 2020-02
    78/100 stars
      Buy from Supplier

    86
    Millipore n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid/product/Millipore
    Average 86 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid - by Bioz Stars, 2020-02
    86/100 stars
      Buy from Supplier

    78
    GE Healthcare n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid Hepes, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 78/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes/product/GE Healthcare
    Average 78 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes - by Bioz Stars, 2020-02
    78/100 stars
      Buy from Supplier

    79
    Fisher Scientific n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid/product/Fisher Scientific
    Average 79 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid - by Bioz Stars, 2020-02
    79/100 stars
      Buy from Supplier

    79
    Nacalai n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid, supplied by Nacalai, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid/product/Nacalai
    Average 79 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid - by Bioz Stars, 2020-02
    79/100 stars
      Buy from Supplier

    79
    Thermo Fisher n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid/product/Thermo Fisher
    Average 79 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid - by Bioz Stars, 2020-02
    79/100 stars
      Buy from Supplier

    82
    Cellgro n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid, supplied by Cellgro, used in various techniques. Bioz Stars score: 82/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid/product/Cellgro
    Average 82 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid - by Bioz Stars, 2020-02
    82/100 stars
      Buy from Supplier

    77
    Atlanta Biologicals sterile hepes n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid buffered rpmi 1640 culture medium
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer <t>(HEPES</t> 20 <t>mmol/L,</t> NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Sterile Hepes N 2 Hydroxyethylpiperazine N 2 Ethanesulfonic Acid Buffered Rpmi 1640 Culture Medium, supplied by Atlanta Biologicals, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sterile hepes n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid buffered rpmi 1640 culture medium/product/Atlanta Biologicals
    Average 77 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    sterile hepes n 2 hydroxyethylpiperazine n 2 ethanesulfonic acid buffered rpmi 1640 culture medium - by Bioz Stars, 2020-02
    77/100 stars
      Buy from Supplier

    90
    Tocris hepes buffer
    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical <t>NPMV</t> fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM <t>HEPES</t> (pH 7.5).
    Hepes Buffer, supplied by Tocris, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes buffer/product/Tocris
    Average 90 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    hepes buffer - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    97
    Amresco hepes
    Ni 2+ and <t>L-cys</t> binding to coptisine (Cop) measured using UV-visible spectroscopy. 16 μM Cop in the absence or presence of different molar equivalents of Ni 2+ (1 mM NiCl 2 ) (A) or L-cys (1 mM) (B) in 20 mM <t>HEPES</t> buffer, pH 7.5. A-F, 0.0 to 320 μM of Ni 2+ ; A-F, 0.0 to 200 μM of L-cys. The inset shows the titration curves for binding of Ni 2+ or L-cys to Cop at 355 nm. Wavelength (nm) is represented on the x -axis and absorption (Abs) is represented on the y -axis.
    Hepes, supplied by Amresco, used in various techniques. Bioz Stars score: 97/100, based on 177 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Amresco
    Average 97 stars, based on 177 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Applichem hepes
    Schematic Workflow for the DTR versus FASP comparison. Four single, deparaffinized murine kidney FFPE tissues were separately prepared with each of the three sample preparation protocol. For the DTR protocol, a buffer containing 0.1% <t>Rapigest</t> in 0.1 M <t>HEPES</t> pH 8 and 1 mM DTT was used for heat-induced antigen retrieval (HIAR) and lysis of the tissue. As RapiGest is compatible with tryptic digestion, direct trypsinization is the key feature of the DTR protocol. RapiGest is later removed by acidifying the sample. Protein concentration is estimated to not overload the C18 stage tips during desalting step and later to inject the same amounts of peptides into the mass spectrometer. The FASP protocol makes use of a buffer containing 4% SDS, 0.1 M HEPES pH 7.5 and 0.05 M DTT. Before digestion the SDS is removed using centrifugal filter units with nominal molecular weight cut offs of either 10,000 or 30,000 Da. After digestion, the peptides are eluted from the filter units and desalted before mass-spectrometry analysis
    Hepes, supplied by Applichem, used in various techniques. Bioz Stars score: 97/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Applichem
    Average 97 stars, based on 170 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    95
    Atlanta Biologicals hepes
    Schematic Workflow for the DTR versus FASP comparison. Four single, deparaffinized murine kidney FFPE tissues were separately prepared with each of the three sample preparation protocol. For the DTR protocol, a buffer containing 0.1% <t>Rapigest</t> in 0.1 M <t>HEPES</t> pH 8 and 1 mM DTT was used for heat-induced antigen retrieval (HIAR) and lysis of the tissue. As RapiGest is compatible with tryptic digestion, direct trypsinization is the key feature of the DTR protocol. RapiGest is later removed by acidifying the sample. Protein concentration is estimated to not overload the C18 stage tips during desalting step and later to inject the same amounts of peptides into the mass spectrometer. The FASP protocol makes use of a buffer containing 4% SDS, 0.1 M HEPES pH 7.5 and 0.05 M DTT. Before digestion the SDS is removed using centrifugal filter units with nominal molecular weight cut offs of either 10,000 or 30,000 Da. After digestion, the peptides are eluted from the filter units and desalted before mass-spectrometry analysis
    Hepes, supplied by Atlanta Biologicals, used in various techniques. Bioz Stars score: 95/100, based on 175 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Atlanta Biologicals
    Average 95 stars, based on 175 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    97
    Avantor hepes
    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml <t>ciprofloxacin</t> in <t>HEPES</t> buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
    Hepes, supplied by Avantor, used in various techniques. Bioz Stars score: 97/100, based on 321 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Avantor
    Average 97 stars, based on 321 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Beyotime hepes
    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml <t>ciprofloxacin</t> in <t>HEPES</t> buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
    Hepes, supplied by Beyotime, used in various techniques. Bioz Stars score: 97/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Beyotime
    Average 97 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Biochrom hepes
    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml <t>ciprofloxacin</t> in <t>HEPES</t> buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
    Hepes, supplied by Biochrom, used in various techniques. Bioz Stars score: 97/100, based on 580 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Biochrom
    Average 97 stars, based on 580 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Biological Industries Inc hepes
    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml <t>ciprofloxacin</t> in <t>HEPES</t> buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
    Hepes, supplied by Biological Industries Inc, used in various techniques. Bioz Stars score: 97/100, based on 330 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Biological Industries Inc
    Average 97 stars, based on 330 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Cambrex hepes
    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml <t>ciprofloxacin</t> in <t>HEPES</t> buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
    Hepes, supplied by Cambrex, used in various techniques. Bioz Stars score: 97/100, based on 254 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Cambrex
    Average 97 stars, based on 254 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Carl Roth GmbH hepes
    Extracellular NETotic DNA is rich in MPO. Representative fluorescence images of NETs induced by CaI (4 μM), PMA (100 nM), or LPS (25 μg/ml) activated in supplement-free <t>RPMI/HEPES.</t> The images show a clear colocalization of MPO (red) with the extracellular DNA-fibers (blue) of released NETs. Scale = 10 μm.
    Hepes, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 97/100, based on 402 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Carl Roth GmbH
    Average 97 stars, based on 402 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Cellgro hepes
    Extracellular NETotic DNA is rich in MPO. Representative fluorescence images of NETs induced by CaI (4 μM), PMA (100 nM), or LPS (25 μg/ml) activated in supplement-free <t>RPMI/HEPES.</t> The images show a clear colocalization of MPO (red) with the extracellular DNA-fibers (blue) of released NETs. Scale = 10 μm.
    Hepes, supplied by Cellgro, used in various techniques. Bioz Stars score: 97/100, based on 838 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Cellgro
    Average 97 stars, based on 838 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    96
    CooperSurgical hepes
    Extracellular NETotic DNA is rich in MPO. Representative fluorescence images of NETs induced by CaI (4 μM), PMA (100 nM), or LPS (25 μg/ml) activated in supplement-free <t>RPMI/HEPES.</t> The images show a clear colocalization of MPO (red) with the extracellular DNA-fibers (blue) of released NETs. Scale = 10 μm.
    Hepes, supplied by CooperSurgical, used in various techniques. Bioz Stars score: 96/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/CooperSurgical
    Average 96 stars, based on 43 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    96/100 stars
      Buy from Supplier

    97
    Corning Life Sciences hepes
    Effect of solution pH on the interaction of Purβ with Acta2 ssDNA. (A) The binding of full-length Purβ (1.0 nM) to PE32-bF ssDNA (0.5 nM) was assessed in assay buffer without MgCl 2 at pH ranging from 7.5 to 12.5. Solid-phase Purβ-PE32-bF complexes were detected by ELISA using rabbit anti-mouse Purβ 210–229 as the primary antibody. Nonspecific background absorbance at 405 nm in control wells without any DNA was subtracted from the signal generated in PE32-bF-coated wells. Background corrected A405 values measured at each pH were normalized by dividing by the mean A405 value determined at pH 7.5 (mean ± SD, n = 4). (B) Effect of solution pH on the thermostability of Purβ. The unfolding of full-length Purβ was evaulated by thermal shift assay at a protein concentration of 2.8 μM in 20 mM <t>HEPES,</t> 150 mM <t>NaCl,</t> 10 mM β-mercaptoethanol adjusted to pH 7.5, 10.5, 11.5, or 12.5.
    Hepes, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 97/100, based on 1105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Corning Life Sciences
    Average 97 stars, based on 1105 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Dojindo Labs hepes
    Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles <t>Mes–KOH,</t> open circles <t>Hepes–KOH,</t> closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b
    Hepes, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 97/100, based on 194 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Dojindo Labs
    Average 97 stars, based on 194 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    93
    EM Science Inc hepes
    Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles <t>Mes–KOH,</t> open circles <t>Hepes–KOH,</t> closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b
    Hepes, supplied by EM Science Inc, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/EM Science Inc
    Average 93 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    93/100 stars
      Buy from Supplier

    97
    EuroClone hepes
    Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles <t>Mes–KOH,</t> open circles <t>Hepes–KOH,</t> closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b
    Hepes, supplied by EuroClone, used in various techniques. Bioz Stars score: 97/100, based on 314 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/EuroClone
    Average 97 stars, based on 314 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    97
    Fisher Scientific hepes
    VIP binding in 100 mM <t>NaCl,</t> 100 mM <t>HEPES</t> at a carrier concentration of 2.4 mg/ml for PGC and 2.0 mg/ml for SSM ( a ) PGC-VIP, ( b ) SSM-VIP. The experiment was done in quadruplicate; free VIP was separated from bound by ultrafiltration, VIP quantitation was
    Hepes, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 97/100, based on 960 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Fisher Scientific
    Average 97 stars, based on 960 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    97/100 stars
      Buy from Supplier

    N/A
    For recreating Screening Suite conditions Kit contents 200 ml 1M HEPES free acid pH 7 5
      Buy from Supplier

    93
    Flow Laboratories hepes
    VIP binding in 100 mM <t>NaCl,</t> 100 mM <t>HEPES</t> at a carrier concentration of 2.4 mg/ml for PGC and 2.0 mg/ml for SSM ( a ) PGC-VIP, ( b ) SSM-VIP. The experiment was done in quadruplicate; free VIP was separated from bound by ultrafiltration, VIP quantitation was
    Hepes, supplied by Flow Laboratories, used in various techniques. Bioz Stars score: 93/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes/product/Flow Laboratories
    Average 93 stars, based on 34 article reviews
    Price from $9.99 to $1999.99
    hepes - by Bioz Stars, 2020-02
    93/100 stars
      Buy from Supplier

    Image Search Results


    (A) Structure of CN-PPV polymer and the absorption (dashed) and emission (solid) spectra of CN-PPV Pdot (black) and CN-PPV Pdot-streptavidin conjugate (CNPPV-Strep, red) in a 0.1% PEG 20 mM HEPES buffer. (B) The absorption and emission spectra of Qdot

    Journal: Macromolecular rapid communications

    Article Title: Ultrasensitive Detection of Proteins on Western Blots with Semiconducting Polymer Dots

    doi: 10.1002/marc.201200809

    Figure Lengend Snippet: (A) Structure of CN-PPV polymer and the absorption (dashed) and emission (solid) spectra of CN-PPV Pdot (black) and CN-PPV Pdot-streptavidin conjugate (CNPPV-Strep, red) in a 0.1% PEG 20 mM HEPES buffer. (B) The absorption and emission spectra of Qdot

    Article Snippet: To conjugate Pdots with streptavidin, to this 5 mL of Pdot solution, we added 100 μL of PEG (5 % w/v in water, MW 3350), 100 μL of 1 M HEPES buffer (pH 7.4), 300 μL of streptavidin (1 mg/mL in 20 mM HEPES buffer), and 50 μL of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC, from Sigma-Aldrich) (10 mg/mL in water).

    Techniques:

    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.

    Article Snippet: The proteins (0.2 mg/mL) were equilibrated in the presence of HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5, Sypro Orange 2.5× (Invitrogen Molecular Probes, http://lifetechnologies.com ).

    Techniques: Ligand Binding Assay, Thermal Shift Assay, Gas Chromatography, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    Specific activity of PMM2 depends on glucose-1,6-bisphosphate concentration. The assay was performed at 32°C in a reaction mixture containing HEPES 20 mmol/L, pH 7.5, MgCl 2 5 mmol/L, NaCl 150 mmol/L, NADP+ 0.25 mmol/L, Glu-1-P (0.04 or 0.60 mmol/L), and yeast glucose 6-phosphate dehydrogenase 10 μg/mL, while Glc-1,6-P 2 was changed in the range 0–80 μmol/L. Enzymes concentrations were 107 nmol/L for wt-PMM2 and 73 nmol/L for F119L-PMM2. The hyperbolic dependence of velocity on the activator concentration was fitted using Michaelis and Menten equation to evaluate EC50.

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Specific activity of PMM2 depends on glucose-1,6-bisphosphate concentration. The assay was performed at 32°C in a reaction mixture containing HEPES 20 mmol/L, pH 7.5, MgCl 2 5 mmol/L, NaCl 150 mmol/L, NADP+ 0.25 mmol/L, Glu-1-P (0.04 or 0.60 mmol/L), and yeast glucose 6-phosphate dehydrogenase 10 μg/mL, while Glc-1,6-P 2 was changed in the range 0–80 μmol/L. Enzymes concentrations were 107 nmol/L for wt-PMM2 and 73 nmol/L for F119L-PMM2. The hyperbolic dependence of velocity on the activator concentration was fitted using Michaelis and Menten equation to evaluate EC50.

    Article Snippet: The proteins (0.2 mg/mL) were equilibrated in the presence of HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5, Sypro Orange 2.5× (Invitrogen Molecular Probes, http://lifetechnologies.com ).

    Techniques: Activity Assay, Concentration Assay, Gas Chromatography

    Long-term stability of F119L-PMM2. F119L-PMM2 (0.027 mmol/L of monomer equivalents) was equilibrated in HEPES 50 mmol/L pH 7.1 containing NaCl 150 mmol/L. Aliquots containing 1.6 μg of protein were taken at known incubation time and diluted immediately to assay the residual activity with Glc-1-P under standard conditions. (A) Results obtained at 37°C in the presence of EDTA 0.1 mmol/L or MgCl 2 5 mmol/L. (B) Results obtained at 44°C in the presence of MgCl 2 5 mmol/L, MgCl 2 5 mmol/L plus Glc-1-P 0.5 mmol/L, MgCl 2 5 mmol/L plus Glc-1-P 0.5 mmol/L and vanadate 0.5 mmol/L or MgCl 2 5 mmol/L plus Glu-1,6-P 2 0.5 mmol/L

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Long-term stability of F119L-PMM2. F119L-PMM2 (0.027 mmol/L of monomer equivalents) was equilibrated in HEPES 50 mmol/L pH 7.1 containing NaCl 150 mmol/L. Aliquots containing 1.6 μg of protein were taken at known incubation time and diluted immediately to assay the residual activity with Glc-1-P under standard conditions. (A) Results obtained at 37°C in the presence of EDTA 0.1 mmol/L or MgCl 2 5 mmol/L. (B) Results obtained at 44°C in the presence of MgCl 2 5 mmol/L, MgCl 2 5 mmol/L plus Glc-1-P 0.5 mmol/L, MgCl 2 5 mmol/L plus Glc-1-P 0.5 mmol/L and vanadate 0.5 mmol/L or MgCl 2 5 mmol/L plus Glu-1,6-P 2 0.5 mmol/L

    Article Snippet: The proteins (0.2 mg/mL) were equilibrated in the presence of HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5, Sypro Orange 2.5× (Invitrogen Molecular Probes, http://lifetechnologies.com ).

    Techniques: Incubation, Activity Assay, Gas Chromatography

    Ligand binding can increase the resistance to proteases of PMM2. Purified wild-type PMM2 and F119L-PMM2 (A) were incubated (0.5 mg/mL) with thermolysin in HEPES 20 mmol/L, NaCl 150 mmol/L, MgCl 2 0.1 mmol/L, pH 7.5 at the indicated protease substrate ratio (w/w) for 1 or 2 h at 37°C before they were analyzed (5 μg of each sample) by SDS-PAGE. Purified F119L-PMM2 (B) was incubated (0.2 mg/mL) with thermolysin in HEPES 20 mmol/L, NaCl 150 mmol/L, MgCl 2 0.1 mmol/L, pH 7.5 at the indicated protease substrate ratio (w/w), in the presence of no ligands, Glc-1,6-P 2 0.5 mmol/L or Glu-6-P 0.5 mmol/L plus vanadate 0.5 mmol/L, for 2 h at 37°C before they were analyzed (2 μg of each sample) by SDS-PAGE. The protein bands were visualized by Coomassie blue staining and the intensity of the bands quantified. The not-digested protein was quantified and expressed as percentage of the starting material (no protease panel A; time 0 panel B).

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Ligand binding can increase the resistance to proteases of PMM2. Purified wild-type PMM2 and F119L-PMM2 (A) were incubated (0.5 mg/mL) with thermolysin in HEPES 20 mmol/L, NaCl 150 mmol/L, MgCl 2 0.1 mmol/L, pH 7.5 at the indicated protease substrate ratio (w/w) for 1 or 2 h at 37°C before they were analyzed (5 μg of each sample) by SDS-PAGE. Purified F119L-PMM2 (B) was incubated (0.2 mg/mL) with thermolysin in HEPES 20 mmol/L, NaCl 150 mmol/L, MgCl 2 0.1 mmol/L, pH 7.5 at the indicated protease substrate ratio (w/w), in the presence of no ligands, Glc-1,6-P 2 0.5 mmol/L or Glu-6-P 0.5 mmol/L plus vanadate 0.5 mmol/L, for 2 h at 37°C before they were analyzed (2 μg of each sample) by SDS-PAGE. The protein bands were visualized by Coomassie blue staining and the intensity of the bands quantified. The not-digested protein was quantified and expressed as percentage of the starting material (no protease panel A; time 0 panel B).

    Article Snippet: The proteins (0.2 mg/mL) were equilibrated in the presence of HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5, Sypro Orange 2.5× (Invitrogen Molecular Probes, http://lifetechnologies.com ).

    Techniques: Ligand Binding Assay, Purification, Incubation, SDS Page, Gas Chromatography, Staining

    Ligand binding affects the quaternary structure of PMM2. Wild-type PMM2 (0.010 mg) and F119L-PMM2 (0.0065 mg) were subjected to size exclusion chromatography on BioSep-SEC-S3000 column equilibrated in HEPES 20 mmol/L pH 7.5, NaCl 150 mmol/L, MgCl 2 5 mmol/L (long dashed line for the wild-type PMM2 or short dashed line for F119L-PMM2) or in the same buffer containing Glc-6-P 0.5 mmol/L and vanadate 0.1 mmol/L (continuous line for wild-type PMM2 or dotted line for F119L-PMM2). The chromatography was run at room temperature at 0.5 mL/min.

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Ligand binding affects the quaternary structure of PMM2. Wild-type PMM2 (0.010 mg) and F119L-PMM2 (0.0065 mg) were subjected to size exclusion chromatography on BioSep-SEC-S3000 column equilibrated in HEPES 20 mmol/L pH 7.5, NaCl 150 mmol/L, MgCl 2 5 mmol/L (long dashed line for the wild-type PMM2 or short dashed line for F119L-PMM2) or in the same buffer containing Glc-6-P 0.5 mmol/L and vanadate 0.1 mmol/L (continuous line for wild-type PMM2 or dotted line for F119L-PMM2). The chromatography was run at room temperature at 0.5 mL/min.

    Article Snippet: The proteins (0.2 mg/mL) were equilibrated in the presence of HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5, Sypro Orange 2.5× (Invitrogen Molecular Probes, http://lifetechnologies.com ).

    Techniques: Ligand Binding Assay, Size-exclusion Chromatography, Gas Chromatography, Chromatography

    Specific activity of F119L-PMM2 depends on enzyme concentration. The assay was performed at 32°C in a reaction mixture containing HEPES 20 mmol/L, pH 7.5, MgCl 2 5 mmol/L, NaCl 150 mmol/L, NADP+ 0.25 mmol/L, Glc-1,6-P 2 0.030 mmol/L, and yeast glucose 6-phosphate dehydrogenase 10 μg/mL. The reaction mixture also contained BSA at 0.5 mg/mL. Three sets of experiments were carried out in the presence of 0.04, 0.16, or 0.6 mmol/L Glc-1-P and the F119L-PMM2 concentration changed in the range 10–240 nmol/L (monomer equivalents).

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Specific activity of F119L-PMM2 depends on enzyme concentration. The assay was performed at 32°C in a reaction mixture containing HEPES 20 mmol/L, pH 7.5, MgCl 2 5 mmol/L, NaCl 150 mmol/L, NADP+ 0.25 mmol/L, Glc-1,6-P 2 0.030 mmol/L, and yeast glucose 6-phosphate dehydrogenase 10 μg/mL. The reaction mixture also contained BSA at 0.5 mg/mL. Three sets of experiments were carried out in the presence of 0.04, 0.16, or 0.6 mmol/L Glc-1-P and the F119L-PMM2 concentration changed in the range 10–240 nmol/L (monomer equivalents).

    Article Snippet: The proteins (0.2 mg/mL) were equilibrated in the presence of HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5, Sypro Orange 2.5× (Invitrogen Molecular Probes, http://lifetechnologies.com ).

    Techniques: Activity Assay, Concentration Assay, Gas Chromatography

    Specific activity of wt-PMM2 changes as a function of protein concentration. The assay was performed at 32°C in a reaction mixture containing HEPES 20 mmol/L, pH 7.5, MgCl 2 5 mmol/L, NaCl 150 mmol/L, NADP+ 0.25 mmol/L, Glc-1,6-P 2 0.030 mmol/L and yeast glucose 6-phosphate dehydrogenase 0.010 mg/mL. The reaction mixture also contained Glc-1-P 0.020 mmol/L and BSA 0.5 mg/mL. The wt-PMM2 concentration changed in the range 2–110 nmol/L (monomer equivalents).

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Specific activity of wt-PMM2 changes as a function of protein concentration. The assay was performed at 32°C in a reaction mixture containing HEPES 20 mmol/L, pH 7.5, MgCl 2 5 mmol/L, NaCl 150 mmol/L, NADP+ 0.25 mmol/L, Glc-1,6-P 2 0.030 mmol/L and yeast glucose 6-phosphate dehydrogenase 0.010 mg/mL. The reaction mixture also contained Glc-1-P 0.020 mmol/L and BSA 0.5 mg/mL. The wt-PMM2 concentration changed in the range 2–110 nmol/L (monomer equivalents).

    Article Snippet: The proteins (0.2 mg/mL) were equilibrated in the presence of HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5, Sypro Orange 2.5× (Invitrogen Molecular Probes, http://lifetechnologies.com ).

    Techniques: Activity Assay, Protein Concentration, Gas Chromatography, Concentration Assay

    Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical NPMV fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM HEPES (pH 7.5).

    Journal: PLoS ONE

    Article Title: Glutamate Uptake Triggers Transporter-Mediated GABA Release from Astrocytes

    doi: 10.1371/journal.pone.0007153

    Figure Lengend Snippet: Glu-induced GABA release is not mediated by vesicular release. (A) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in the presence of glutamate receptor antagonists (n = 7; P = 0.014; 128.5±13.4% of pre-stimulus control). (B) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 and Glu receptor antagonists in rat cerebrocortical NPMV fractions (n = 4; P = 0.82) (C) Elevation of [GABA] o in acute rat hippocampal slices in control conditions (n = 6) and in low-[Ca 2+ ]/high-[Mg 2+ ] buffer (n = 5; P = 0.91; 148.3±15.5% of pre-stimulus control). (D) Cytosolic GABA level as determined by [ 3 H]GABA uptake in the presence of 100 µM NNC-711 in low-[Ca 2+ ] buffer in rat cerebrocortical NPMV fractions (n = 3; P = 0.49). Low-[Ca 2+ ] buffer contained 145 mM NaCl, 5 mM KCl, 20 mM MgCl 2 , 10 mM glucose and 20 mM HEPES (pH 7.5).

    Article Snippet: Uptake studies were performed as follows: aliquots of NPMV fractions in HEPES buffer were preincubated with test compounds in the presence of NNC-711 (100 µM, Tocris) at 30°C for 10 min.

    Techniques:

    Ni 2+ and L-cys binding to coptisine (Cop) measured using UV-visible spectroscopy. 16 μM Cop in the absence or presence of different molar equivalents of Ni 2+ (1 mM NiCl 2 ) (A) or L-cys (1 mM) (B) in 20 mM HEPES buffer, pH 7.5. A-F, 0.0 to 320 μM of Ni 2+ ; A-F, 0.0 to 200 μM of L-cys. The inset shows the titration curves for binding of Ni 2+ or L-cys to Cop at 355 nm. Wavelength (nm) is represented on the x -axis and absorption (Abs) is represented on the y -axis.

    Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

    Article Title: Coptisine-induced inhibition of Helicobacter pylori: elucidation of specific mechanisms by probing urease active site and its maturation process

    doi: 10.1080/14756366.2018.1501044

    Figure Lengend Snippet: Ni 2+ and L-cys binding to coptisine (Cop) measured using UV-visible spectroscopy. 16 μM Cop in the absence or presence of different molar equivalents of Ni 2+ (1 mM NiCl 2 ) (A) or L-cys (1 mM) (B) in 20 mM HEPES buffer, pH 7.5. A-F, 0.0 to 320 μM of Ni 2+ ; A-F, 0.0 to 200 μM of L-cys. The inset shows the titration curves for binding of Ni 2+ or L-cys to Cop at 355 nm. Wavelength (nm) is represented on the x -axis and absorption (Abs) is represented on the y -axis.

    Article Snippet: Acetohydroxamic acid (AHA; C2 H5 NO2 , purity: 98%), jack bean urease (JBU; type III with activity of 31.66 U/mg solid), urea, dithiothreitol (DTT), L-cysteine (L-cys), glutathione (GSH), sodium fluoride (NaF), and HEPES (Amresco > 99%) were purchased from Sigma Aldrich (Steineheim, Germany).

    Techniques: Binding Assay, Spectroscopy, Titration

    Schematic Workflow for the DTR versus FASP comparison. Four single, deparaffinized murine kidney FFPE tissues were separately prepared with each of the three sample preparation protocol. For the DTR protocol, a buffer containing 0.1% Rapigest in 0.1 M HEPES pH 8 and 1 mM DTT was used for heat-induced antigen retrieval (HIAR) and lysis of the tissue. As RapiGest is compatible with tryptic digestion, direct trypsinization is the key feature of the DTR protocol. RapiGest is later removed by acidifying the sample. Protein concentration is estimated to not overload the C18 stage tips during desalting step and later to inject the same amounts of peptides into the mass spectrometer. The FASP protocol makes use of a buffer containing 4% SDS, 0.1 M HEPES pH 7.5 and 0.05 M DTT. Before digestion the SDS is removed using centrifugal filter units with nominal molecular weight cut offs of either 10,000 or 30,000 Da. After digestion, the peptides are eluted from the filter units and desalted before mass-spectrometry analysis

    Journal: Clinical Proteomics

    Article Title: Reproducible proteomics sample preparation for single FFPE tissue slices using acid-labile surfactant and direct trypsinization

    doi: 10.1186/s12014-018-9188-y

    Figure Lengend Snippet: Schematic Workflow for the DTR versus FASP comparison. Four single, deparaffinized murine kidney FFPE tissues were separately prepared with each of the three sample preparation protocol. For the DTR protocol, a buffer containing 0.1% Rapigest in 0.1 M HEPES pH 8 and 1 mM DTT was used for heat-induced antigen retrieval (HIAR) and lysis of the tissue. As RapiGest is compatible with tryptic digestion, direct trypsinization is the key feature of the DTR protocol. RapiGest is later removed by acidifying the sample. Protein concentration is estimated to not overload the C18 stage tips during desalting step and later to inject the same amounts of peptides into the mass spectrometer. The FASP protocol makes use of a buffer containing 4% SDS, 0.1 M HEPES pH 7.5 and 0.05 M DTT. Before digestion the SDS is removed using centrifugal filter units with nominal molecular weight cut offs of either 10,000 or 30,000 Da. After digestion, the peptides are eluted from the filter units and desalted before mass-spectrometry analysis

    Article Snippet: 100 µl (200 µl for the tonsil samples) of an aqueous buffer containing 0.1% RapiGest SF (Waters, Milford, MA, USA), 0.1 M HEPES pH 8 (AppliChem, Darmstadt, Germany) and 1 mM dithiothreitol (DTT) (AppliChem, Darmstadt, Germany) were added to each reaction tube, containing a single tissue slice.

    Techniques: Formalin-fixed Paraffin-Embedded, Sample Prep, Lysis, Protein Concentration, Mass Spectrometry, Molecular Weight

    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.

    Journal: Pharmaceutical Research

    Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug

    doi: 10.1007/s11095-016-2002-5

    Figure Lengend Snippet: Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.

    Article Snippet: The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)).

    Techniques: Incubation

    Evaluation of the effect of freeze-thaw in liquid nitrogen on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and 0, 0.05 or 0.1% polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control prior to freeze-thaw ( blue diamonds ) or after freeze-thaw ( red circles ), CFI containing 0.05% polysorbate 20 after freeze-thaw ( green diamonds ), CFI containing 0.1% polysorbate 20 after freeze-thaw ( yellow triangles ). Duplicate samples were analyzed at each time point. Two simulated curves were generated by adjusting the release from the CFI sample without polysorbate 20 after freeze-thaw by assuming the addition of either 10% free drug ( green dashed line ) or 20% free drug ( yellow dashed line ) and normalizing the total drug release to 100%.

    Journal: Pharmaceutical Research

    Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug

    doi: 10.1007/s11095-016-2002-5

    Figure Lengend Snippet: Evaluation of the effect of freeze-thaw in liquid nitrogen on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and 0, 0.05 or 0.1% polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control prior to freeze-thaw ( blue diamonds ) or after freeze-thaw ( red circles ), CFI containing 0.05% polysorbate 20 after freeze-thaw ( green diamonds ), CFI containing 0.1% polysorbate 20 after freeze-thaw ( yellow triangles ). Duplicate samples were analyzed at each time point. Two simulated curves were generated by adjusting the release from the CFI sample without polysorbate 20 after freeze-thaw by assuming the addition of either 10% free drug ( green dashed line ) or 20% free drug ( yellow dashed line ) and normalizing the total drug release to 100%.

    Article Snippet: The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)).

    Techniques: Incubation, Generated

    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and Brij 30 before ( a ) and after ( b ) freeze-thaw. The 12.5 mg/ml CFI formulations were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze thaw, blue diamonds ), CFI containing 0.05% Brij 30 ( red squares ), CFI containing 0.1% Brij 30 ( green triangles ), CFI containing 0.2% Brij 30 ( orange circles ), and CFI containing 0.3% Brij 30 ( purple squares ). Duplicate samples were analyzed at each time point.

    Journal: Pharmaceutical Research

    Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug

    doi: 10.1007/s11095-016-2002-5

    Figure Lengend Snippet: Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and Brij 30 before ( a ) and after ( b ) freeze-thaw. The 12.5 mg/ml CFI formulations were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze thaw, blue diamonds ), CFI containing 0.05% Brij 30 ( red squares ), CFI containing 0.1% Brij 30 ( green triangles ), CFI containing 0.2% Brij 30 ( orange circles ), and CFI containing 0.3% Brij 30 ( purple squares ). Duplicate samples were analyzed at each time point.

    Article Snippet: The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)).

    Techniques: Incubation

    Extracellular NETotic DNA is rich in MPO. Representative fluorescence images of NETs induced by CaI (4 μM), PMA (100 nM), or LPS (25 μg/ml) activated in supplement-free RPMI/HEPES. The images show a clear colocalization of MPO (red) with the extracellular DNA-fibers (blue) of released NETs. Scale = 10 μm.

    Journal: Frontiers in Immunology

    Article Title: Serum and Serum Albumin Inhibit in vitro Formation of Neutrophil Extracellular Traps (NETs)

    doi: 10.3389/fimmu.2019.00012

    Figure Lengend Snippet: Extracellular NETotic DNA is rich in MPO. Representative fluorescence images of NETs induced by CaI (4 μM), PMA (100 nM), or LPS (25 μg/ml) activated in supplement-free RPMI/HEPES. The images show a clear colocalization of MPO (red) with the extracellular DNA-fibers (blue) of released NETs. Scale = 10 μm.

    Article Snippet: Freshly isolated human or murine neutrophils were counted and suspended in RPMI 1640 (Lonza) containing 10 mM HEPES (Roth) (RPMI/HEPES) and 0.5% HSA (Sigma-Aldrich), 0.5%/1%/2% FCS (Biochrom GmbH, Merck Millipore) or 0.5% BSA (Roth), respectively.

    Techniques: Fluorescence

    Influence of serum and serum albumin supplements on NET formation of murine neutrophils. (A) Representative fluorescence images of nuclei of murine neutrophils (Hoechst) after stimulation with CaI (4 μM), PMA (100 nM) or LPS (25 μg/ml) for 180 min, respectively. NET formation of neutrophils was studied in RPMI/ HEPES without supplements and RPMI/ HEPES supplemented with 0.5% BSA or 2% hiFCS, as indicated. Chromatin decondensation is only inducible in RPMI/ HEPES (white arrow heads). Scale = 50 μm. (B) Percentage of decondensed nuclei/ NETs after stimulation with PMA (100 nM), CaI (4 μM), or LPS (10, 25, or 100 μg/ml) for 180 min. Murine neutrophils were studied in RPMI/ HEPES and RPMI/ HEPES supplemented with 0.5% BSA or 2% hiFCS. Addition of 0.5% BSA or 2% hiFCS to RPMI/ HEPES, inhibits chromatin decondensation induced by PMA, CaI and LPS completely. Error bars = mean ± SEM. ns = not significant. **** p

    Journal: Frontiers in Immunology

    Article Title: Serum and Serum Albumin Inhibit in vitro Formation of Neutrophil Extracellular Traps (NETs)

    doi: 10.3389/fimmu.2019.00012

    Figure Lengend Snippet: Influence of serum and serum albumin supplements on NET formation of murine neutrophils. (A) Representative fluorescence images of nuclei of murine neutrophils (Hoechst) after stimulation with CaI (4 μM), PMA (100 nM) or LPS (25 μg/ml) for 180 min, respectively. NET formation of neutrophils was studied in RPMI/ HEPES without supplements and RPMI/ HEPES supplemented with 0.5% BSA or 2% hiFCS, as indicated. Chromatin decondensation is only inducible in RPMI/ HEPES (white arrow heads). Scale = 50 μm. (B) Percentage of decondensed nuclei/ NETs after stimulation with PMA (100 nM), CaI (4 μM), or LPS (10, 25, or 100 μg/ml) for 180 min. Murine neutrophils were studied in RPMI/ HEPES and RPMI/ HEPES supplemented with 0.5% BSA or 2% hiFCS. Addition of 0.5% BSA or 2% hiFCS to RPMI/ HEPES, inhibits chromatin decondensation induced by PMA, CaI and LPS completely. Error bars = mean ± SEM. ns = not significant. **** p

    Article Snippet: Freshly isolated human or murine neutrophils were counted and suspended in RPMI 1640 (Lonza) containing 10 mM HEPES (Roth) (RPMI/HEPES) and 0.5% HSA (Sigma-Aldrich), 0.5%/1%/2% FCS (Biochrom GmbH, Merck Millipore) or 0.5% BSA (Roth), respectively.

    Techniques: Fluorescence

    Influence of serum and serum albumin supplements on NET formation of human neutrophils. (A) Representative fluorescence images of human neutrophils (chromatin stained by Hoechst) after stimulation with CaI (4 μM), PMA (100 nM), or LPS (25 μg/ml) for 180 min, respectively. NET formation of neutrophils was studied in RPMI 1,640 with 10 mM HEPES (RPMI/HEPES), RPMI/ HEPES + 0.5% human serum albumin (HSA) or + 1% heat inactivated (56°C) fetal calf serum (hiFCS). Chromatin decondensation induced by PMA is clearly visible with all three culture conditions, while LPS or CaI only cause NET formation in BSA- and HSA-free RPMI/HEPES. Scale = 50 μm. (B) Neutrophils were stimulated to undergo NET formation with CaI (4 μM), PMA (100 nM) or LPS (10, 25, or 100 μg/ml), respectively. Both, HSA and BSA inhibit CaI and LPS-induced formation of NETs (determined as percentage of decondensed nuclei/NETs of total neutrophils). NETosis stimulated by PMA is independent of serum albumin addition. Error bars = mean ± SEM. ns, not significant. * p

    Journal: Frontiers in Immunology

    Article Title: Serum and Serum Albumin Inhibit in vitro Formation of Neutrophil Extracellular Traps (NETs)

    doi: 10.3389/fimmu.2019.00012

    Figure Lengend Snippet: Influence of serum and serum albumin supplements on NET formation of human neutrophils. (A) Representative fluorescence images of human neutrophils (chromatin stained by Hoechst) after stimulation with CaI (4 μM), PMA (100 nM), or LPS (25 μg/ml) for 180 min, respectively. NET formation of neutrophils was studied in RPMI 1,640 with 10 mM HEPES (RPMI/HEPES), RPMI/ HEPES + 0.5% human serum albumin (HSA) or + 1% heat inactivated (56°C) fetal calf serum (hiFCS). Chromatin decondensation induced by PMA is clearly visible with all three culture conditions, while LPS or CaI only cause NET formation in BSA- and HSA-free RPMI/HEPES. Scale = 50 μm. (B) Neutrophils were stimulated to undergo NET formation with CaI (4 μM), PMA (100 nM) or LPS (10, 25, or 100 μg/ml), respectively. Both, HSA and BSA inhibit CaI and LPS-induced formation of NETs (determined as percentage of decondensed nuclei/NETs of total neutrophils). NETosis stimulated by PMA is independent of serum albumin addition. Error bars = mean ± SEM. ns, not significant. * p

    Article Snippet: Freshly isolated human or murine neutrophils were counted and suspended in RPMI 1640 (Lonza) containing 10 mM HEPES (Roth) (RPMI/HEPES) and 0.5% HSA (Sigma-Aldrich), 0.5%/1%/2% FCS (Biochrom GmbH, Merck Millipore) or 0.5% BSA (Roth), respectively.

    Techniques: Fluorescence, Staining, Albumin Addition

    Effect of solution pH on the interaction of Purβ with Acta2 ssDNA. (A) The binding of full-length Purβ (1.0 nM) to PE32-bF ssDNA (0.5 nM) was assessed in assay buffer without MgCl 2 at pH ranging from 7.5 to 12.5. Solid-phase Purβ-PE32-bF complexes were detected by ELISA using rabbit anti-mouse Purβ 210–229 as the primary antibody. Nonspecific background absorbance at 405 nm in control wells without any DNA was subtracted from the signal generated in PE32-bF-coated wells. Background corrected A405 values measured at each pH were normalized by dividing by the mean A405 value determined at pH 7.5 (mean ± SD, n = 4). (B) Effect of solution pH on the thermostability of Purβ. The unfolding of full-length Purβ was evaulated by thermal shift assay at a protein concentration of 2.8 μM in 20 mM HEPES, 150 mM NaCl, 10 mM β-mercaptoethanol adjusted to pH 7.5, 10.5, 11.5, or 12.5.

    Journal: Biochemistry

    Article Title: Electrostatic and Hydrophobic Interactions Mediate Single-Stranded DNA Recognition and Acta2 Repression by Purine-Rich Element Binding Protein B

    doi: 10.1021/acs.biochem.6b00006

    Figure Lengend Snippet: Effect of solution pH on the interaction of Purβ with Acta2 ssDNA. (A) The binding of full-length Purβ (1.0 nM) to PE32-bF ssDNA (0.5 nM) was assessed in assay buffer without MgCl 2 at pH ranging from 7.5 to 12.5. Solid-phase Purβ-PE32-bF complexes were detected by ELISA using rabbit anti-mouse Purβ 210–229 as the primary antibody. Nonspecific background absorbance at 405 nm in control wells without any DNA was subtracted from the signal generated in PE32-bF-coated wells. Background corrected A405 values measured at each pH were normalized by dividing by the mean A405 value determined at pH 7.5 (mean ± SD, n = 4). (B) Effect of solution pH on the thermostability of Purβ. The unfolding of full-length Purβ was evaulated by thermal shift assay at a protein concentration of 2.8 μM in 20 mM HEPES, 150 mM NaCl, 10 mM β-mercaptoethanol adjusted to pH 7.5, 10.5, 11.5, or 12.5.

    Article Snippet: Briefly, NHis-tagged MSY1 was diluted to 100 nM in coating buffer consisting of 20 mM HEPES, 150 mM NaCl, 1.5 mM MgCl2 -6H2 O pH 7.5 with 5.0 μg/ml BSA and then applied to microtiter wells (Costar® EIA/RIA plate, 96 Well Easy Wash™, Certified High Binding, Corning, Inc.).

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Generated, Thermal Shift Assay, Protein Concentration

    Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles Mes–KOH, open circles Hepes–KOH, closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b

    Journal: Planta

    Article Title: Biosynthesis of the carbohydrate moieties of arabinogalactan proteins by membrane-bound ?-glucuronosyltransferases from radish primary roots

    doi: 10.1007/s00425-013-1959-0

    Figure Lengend Snippet: Activity of β-GlcATs in microsomes from radish primary roots. a The microsomes (protein, 50 μg) were incubated under standard assay conditions with β-(1 → 3)-galactan as exogenous acceptor ( closed circles ) and with endogenous acceptors ( open circles ) for varying times up to 240 min. After 240 min, the amount of [ 14 C]GlcA transferred to β-(1 → 3)-galactan was about 0.6 % of initial UDP-[ 14 C]GlcA. The data are the mean + SE of quadruple experiments. b Relationship between enzyme activity and protein concentration. Various amounts of microsomal protein were incubated under standard conditions for 60 min. c Effects of pH on enzyme activity. Activity–pH curves result from experiments with buffers (80 mM) having different pH values under standard conditions for 120 min. Closed circles Mes–KOH, open circles Hepes–KOH, closed triangles acetate. d Activity–temperature curves result from incubations at different temperatures under standard assay conditions for 120 min. Note that the activity plotted for β-(1 → 3)-galactan includes that for endogenous acceptors in a and b

    Article Snippet: EGTA, Hepes, and Mes were from Dojindo Laboratories (Kumamoto, Japan). β-(1 → 3)- and β-(1 → 6)-Galactobioses, and -trioses were prepared from larch wood AG by partial acid hydrolysis (Aspinall et al. ).

    Techniques: Activity Assay, Incubation, Protein Concentration

    VIP binding in 100 mM NaCl, 100 mM HEPES at a carrier concentration of 2.4 mg/ml for PGC and 2.0 mg/ml for SSM ( a ) PGC-VIP, ( b ) SSM-VIP. The experiment was done in quadruplicate; free VIP was separated from bound by ultrafiltration, VIP quantitation was

    Journal: Pharmaceutical research

    Article Title: Protected Graft Copolymer Excipient Leads to a Higher Acute Maximum Tolerated Dose and Extends Residence Time of Vasoactive Intestinal Peptide Significantly Better than Sterically Stabilized Micelles

    doi: 10.1007/s11095-012-0904-4

    Figure Lengend Snippet: VIP binding in 100 mM NaCl, 100 mM HEPES at a carrier concentration of 2.4 mg/ml for PGC and 2.0 mg/ml for SSM ( a ) PGC-VIP, ( b ) SSM-VIP. The experiment was done in quadruplicate; free VIP was separated from bound by ultrafiltration, VIP quantitation was

    Article Snippet: VIP was mixed with a fixed amount of PGC (0.6 mg at 2.4 mg/ml final concentration) or SSM carriers (0.5 mg at 2 mg/ml final concentration) at concentrations between 72 μM and 188 μM in the PGC mixtures and between 120 μM and 240 μM in the SSM mixtures in an aqueous solution of 100 mM NaCl (Fisher Scientific, Waltham MA) and 100 mM HEPES (Fisher Scientific, Waltham MA) in quadruplicate.

    Techniques: Binding Assay, Concentration Assay, Pyrolysis Gas Chromatography, Quantitation Assay