proteinase k conjugated to agarose beads  (Millipore)


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    Structured Review

    Millipore proteinase k conjugated to agarose beads
    Secreted factor(s) have protein qualities. Whole cell protein extracts from HEp-2 cells analyzed by immunoblotting showed that ( A ) Incubation with heat treated (100°C, 0.5h) EHEC CS did not suppress IFNγ mediated Stat-1 tyrosine phosphorylation, and ( B ) incubation with EHEC CS (6h) pre-treated with <t>proteinase</t> K conjugated to agarose beads (0 – 1,000mg/ml, 37°C, 1h) did not suppress IFNγ induced tyrosine phosphorylation of Stat-1 (n = 3, one-way ANOVA, * p
    Proteinase K Conjugated To Agarose Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/proteinase k conjugated to agarose beads/product/Millipore
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    proteinase k conjugated to agarose beads - by Bioz Stars, 2020-04
    91/100 stars

    Related Products / Commonly Used Together

    ecpcm
    ehec
    proteinase k

    Images

    1) Product Images from "Identifying Mechanisms by Which Escherichia coli O157:H7 Subverts Interferon-? Mediated Signal Transducer and Activator of Transcription-1 Activation"

    Article Title: Identifying Mechanisms by Which Escherichia coli O157:H7 Subverts Interferon-? Mediated Signal Transducer and Activator of Transcription-1 Activation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0030145

    Secreted factor(s) have protein qualities. Whole cell protein extracts from HEp-2 cells analyzed by immunoblotting showed that ( A ) Incubation with heat treated (100°C, 0.5h) EHEC CS did not suppress IFNγ mediated Stat-1 tyrosine phosphorylation, and ( B ) incubation with EHEC CS (6h) pre-treated with proteinase K conjugated to agarose beads (0 – 1,000mg/ml, 37°C, 1h) did not suppress IFNγ induced tyrosine phosphorylation of Stat-1 (n = 3, one-way ANOVA, * p
    Figure Legend Snippet: Secreted factor(s) have protein qualities. Whole cell protein extracts from HEp-2 cells analyzed by immunoblotting showed that ( A ) Incubation with heat treated (100°C, 0.5h) EHEC CS did not suppress IFNγ mediated Stat-1 tyrosine phosphorylation, and ( B ) incubation with EHEC CS (6h) pre-treated with proteinase K conjugated to agarose beads (0 – 1,000mg/ml, 37°C, 1h) did not suppress IFNγ induced tyrosine phosphorylation of Stat-1 (n = 3, one-way ANOVA, * p

    Techniques Used: Incubation

    2) Product Images from "A Secreted Factor Coordinates Environmental Quality with Bacillus Development"

    Article Title: A Secreted Factor Coordinates Environmental Quality with Bacillus Development

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0144168

    FacX is resistant to boiling, but sensitive to Proteinase K. (A) Conditioned media was boiled for 15 min and utilized in the sporulation assay ( Fig 2A ) with strain MF1913 (P hy-spank - kinA ). (B) Conditioned media was treated with proteinase K and utilized in the sporulation assay with strain MF1913 (P hy-spank - kinA ). When indicated, KinA was induced with IPTG (20 μM). Cells were grown for 2 hrs at 37°C before image capture. Membranes were stained with TMA.
    Figure Legend Snippet: FacX is resistant to boiling, but sensitive to Proteinase K. (A) Conditioned media was boiled for 15 min and utilized in the sporulation assay ( Fig 2A ) with strain MF1913 (P hy-spank - kinA ). (B) Conditioned media was treated with proteinase K and utilized in the sporulation assay with strain MF1913 (P hy-spank - kinA ). When indicated, KinA was induced with IPTG (20 μM). Cells were grown for 2 hrs at 37°C before image capture. Membranes were stained with TMA.

    Techniques Used: Staining

    3) Product Images from "A novel endothelial-derived anti-inflammatory activity significantly inhibits spontaneous choroidal neovascularisation in a mouse model"

    Article Title: A novel endothelial-derived anti-inflammatory activity significantly inhibits spontaneous choroidal neovascularisation in a mouse model

    Journal: Vascular Cell

    doi: 10.1186/s13221-016-0036-4

    a Dose-dependent anti-inflammatory effects of ECPCM. HAEC were treated for 2 h with TNFα in collection medium, ECPCM, or serial dilutions (1:2) of ECPCM in collection medium. The graph shows the percentage of expression of E-selectin and VCAM-1 relative to that observed for TNFα in collection medium, as determined by real-time PCR analysis. Data = mean ± SEM. b ECPCM anti-inflammatory activity is not affected by proteinase K or RNase treatment. Gene expression of E-selectin and VCAM-1 was analysed by real-time PCR after treatment for 2 h with TNFα. HAEC were treated with collection medium, ECPCM or ECPCM previously treated with proteinase K or RNase. p value: *
    Figure Legend Snippet: a Dose-dependent anti-inflammatory effects of ECPCM. HAEC were treated for 2 h with TNFα in collection medium, ECPCM, or serial dilutions (1:2) of ECPCM in collection medium. The graph shows the percentage of expression of E-selectin and VCAM-1 relative to that observed for TNFα in collection medium, as determined by real-time PCR analysis. Data = mean ± SEM. b ECPCM anti-inflammatory activity is not affected by proteinase K or RNase treatment. Gene expression of E-selectin and VCAM-1 was analysed by real-time PCR after treatment for 2 h with TNFα. HAEC were treated with collection medium, ECPCM or ECPCM previously treated with proteinase K or RNase. p value: *

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Activity Assay

    Related Articles

    Centrifugation:

    Article Title: Identifying Mechanisms by Which Escherichia coli O157:H7 Subverts Interferon-? Mediated Signal Transducer and Activator of Transcription-1 Activation
    Article Snippet: Proteinase K and heat inactivation treatment of culture supernatants Bacterial culture supernatants from EHEC were incubated with proteinase K conjugated to agarose beads (10 to 1000 µg/ml, 1h shaking, 37°C) (Sigma Aldrich, Oakville, Ontario, Canada). .. After incubation, agarose beads were removed from the solution by centrifugation (3,000g, 1 min) before incubation with HEp-2 cells.

    Article Title: Enterohemorrhagic Escherichia coli O157:H7 Shiga Toxins Inhibit Gamma Interferon-Mediated Cellular Activation
    Article Snippet: Bacterial culture supernatants from EHEC were incubated with proteinase K conjugated to agarose beads (10 to 1,000 μg/ml, 1 h, shaking, 37°C) (Sigma-Aldrich, Oakville, Ontario, Canada). .. After incubation, agarose beads were removed from the solution by centrifugation (3,000 × g , 1 min) before incubation with HEp-2 cells.

    Article Title: A novel endothelial-derived anti-inflammatory activity significantly inhibits spontaneous choroidal neovascularisation in a mouse model
    Article Snippet: .. To determine if the active molecule was a protein 1 ml of ECPCM was heated at 95 °C for 15 min and approximately 0.2 units (5.5 mg) of proteinase K conjugated to agarose beads (Sigma, St. Louis, MO) were added, followed by incubation at 55 °C for 16 h. The enzyme was removed from the medium by centrifugation at 2660 × g for 10 min at 4 °C. .. For the RNase A/T1 (Fermentas, Sunderland, UK) treatment 1 ml of ECPCM was incubated with 20 μl of the enzyme mix at 37 °C for 1 h. After each treatment the ECPCM was cooled on ice and then stored at 4 °C until assayed.

    Transferring:

    Article Title: A Secreted Factor Coordinates Environmental Quality with Bacillus Development
    Article Snippet: Heat treatment, proteinase K treatment and dialysis of conditioned media Heat treatment of conditioned CH media was carried out by transferring the conditioned media to a glass tube and submerging the glass tube in a bath of boiling water for 15 min. .. To treat the conditioned CH media with protease, 30 mg of proteinase K conjugated to agarose beads (Sigma) was hydrated in 3 ml ddH2 0 for 20 min at room temperature, and then centrifuged at 1,292 x g for 2 min. After removing the supernatant, the beads were washed in 3 ml ddH2 0, and pelleted by centrifuging at 1,292 x g for 2 min.

    Negative Control:

    Article Title: Identifying Mechanisms by Which Escherichia coli O157:H7 Subverts Interferon-? Mediated Signal Transducer and Activator of Transcription-1 Activation
    Article Snippet: Proteinase K and heat inactivation treatment of culture supernatants Bacterial culture supernatants from EHEC were incubated with proteinase K conjugated to agarose beads (10 to 1000 µg/ml, 1h shaking, 37°C) (Sigma Aldrich, Oakville, Ontario, Canada). .. Culture supernatants incubated with agarose beads and pre-incubated with bovine serum albumin (5% BSA) were used as a negative control.

    Article Title: Enterohemorrhagic Escherichia coli O157:H7 Shiga Toxins Inhibit Gamma Interferon-Mediated Cellular Activation
    Article Snippet: Bacterial culture supernatants from EHEC were incubated with proteinase K conjugated to agarose beads (10 to 1,000 μg/ml, 1 h, shaking, 37°C) (Sigma-Aldrich, Oakville, Ontario, Canada). .. Culture supernatants incubated with agarose beads and preincubated with 5% bovine serum albumin (BSA) were used as a negative control.

    Enzyme-linked Immunosorbent Assay:

    Article Title: A novel endothelial-derived anti-inflammatory activity significantly inhibits spontaneous choroidal neovascularisation in a mouse model
    Article Snippet: To determine if the active molecule was a protein 1 ml of ECPCM was heated at 95 °C for 15 min and approximately 0.2 units (5.5 mg) of proteinase K conjugated to agarose beads (Sigma, St. Louis, MO) were added, followed by incubation at 55 °C for 16 h. The enzyme was removed from the medium by centrifugation at 2660 × g for 10 min at 4 °C. .. ELISA kits for TGF-β1, interleukin (IL)-10, cyclic AMP (cAMP) (R & D Systems, Abingdon, UK) and prostaglandin I2 (PGI2 ) (MyBioSource, Upper Heyford, UK) were used, according to the manufacturer’s instructions, to determine the concentration of the molecules in collection medium and ECPCM.

    Concentration Assay:

    Article Title: A novel endothelial-derived anti-inflammatory activity significantly inhibits spontaneous choroidal neovascularisation in a mouse model
    Article Snippet: To determine if the active molecule was a protein 1 ml of ECPCM was heated at 95 °C for 15 min and approximately 0.2 units (5.5 mg) of proteinase K conjugated to agarose beads (Sigma, St. Louis, MO) were added, followed by incubation at 55 °C for 16 h. The enzyme was removed from the medium by centrifugation at 2660 × g for 10 min at 4 °C. .. ELISA kits for TGF-β1, interleukin (IL)-10, cyclic AMP (cAMP) (R & D Systems, Abingdon, UK) and prostaglandin I2 (PGI2 ) (MyBioSource, Upper Heyford, UK) were used, according to the manufacturer’s instructions, to determine the concentration of the molecules in collection medium and ECPCM.

    Incubation:

    Article Title: Identifying Mechanisms by Which Escherichia coli O157:H7 Subverts Interferon-? Mediated Signal Transducer and Activator of Transcription-1 Activation
    Article Snippet: .. Proteinase K and heat inactivation treatment of culture supernatants Bacterial culture supernatants from EHEC were incubated with proteinase K conjugated to agarose beads (10 to 1000 µg/ml, 1h shaking, 37°C) (Sigma Aldrich, Oakville, Ontario, Canada). .. Culture supernatants incubated with agarose beads and pre-incubated with bovine serum albumin (5% BSA) were used as a negative control.

    Article Title: Enterohemorrhagic Escherichia coli O157:H7 Shiga Toxins Inhibit Gamma Interferon-Mediated Cellular Activation
    Article Snippet: .. Bacterial culture supernatants from EHEC were incubated with proteinase K conjugated to agarose beads (10 to 1,000 μg/ml, 1 h, shaking, 37°C) (Sigma-Aldrich, Oakville, Ontario, Canada). .. Culture supernatants incubated with agarose beads and preincubated with 5% bovine serum albumin (BSA) were used as a negative control.

    Article Title: A novel endothelial-derived anti-inflammatory activity significantly inhibits spontaneous choroidal neovascularisation in a mouse model
    Article Snippet: .. To determine if the active molecule was a protein 1 ml of ECPCM was heated at 95 °C for 15 min and approximately 0.2 units (5.5 mg) of proteinase K conjugated to agarose beads (Sigma, St. Louis, MO) were added, followed by incubation at 55 °C for 16 h. The enzyme was removed from the medium by centrifugation at 2660 × g for 10 min at 4 °C. .. For the RNase A/T1 (Fermentas, Sunderland, UK) treatment 1 ml of ECPCM was incubated with 20 μl of the enzyme mix at 37 °C for 1 h. After each treatment the ECPCM was cooled on ice and then stored at 4 °C until assayed.

    Software:

    Article Title: A novel endothelial-derived anti-inflammatory activity significantly inhibits spontaneous choroidal neovascularisation in a mouse model
    Article Snippet: GraphPad Prism software was used for non-linear regression analysis of the data for IC50 determination. .. To determine if the active molecule was a protein 1 ml of ECPCM was heated at 95 °C for 15 min and approximately 0.2 units (5.5 mg) of proteinase K conjugated to agarose beads (Sigma, St. Louis, MO) were added, followed by incubation at 55 °C for 16 h. The enzyme was removed from the medium by centrifugation at 2660 × g for 10 min at 4 °C.