tpck trypsin  (Worthington Biochemical)


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    Name:
    Trypsin
    Description:
    Supplied as dialyzed and lyophilized powder
    Catalog Number:
    ls003702
    Price:
    23
    Source:
    Bovine Pancreas
    Size:
    100 mg
    Cas Number:
    9002.07.7
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    Structured Review

    Worthington Biochemical tpck trypsin
    Replication of IAV in pAECs. (A) Primary pAECs were infected with the indicated virus at MOI of 0.01 without exogenous trypsin and the supernatants were collected at 0, 4, 8, 12, 24, 48, 72, and 96 h p.i.. Virus titers were determined by TCID 50 assay in <t>MDCK</t> cells in the presence of 0.5 μg/ml <t>TPCK-trypsin.</t> (B) Titer of IAV pH1N1 CA/09 and H1N1 IL/08 in the culture supernatant of ipAECs 72 h p.i in the presence and absence of exogenous TPCK-trypsin. Data are average of 2 independent experiments (n = 2).
    Supplied as dialyzed and lyophilized powder
    https://www.bioz.com/result/tpck trypsin/product/Worthington Biochemical
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tpck trypsin - by Bioz Stars, 2021-03
    99/100 stars

    Images

    1) Product Images from "Differential Induction of Type I and Type III Interferons by Swine and Human Origin H1N1 Influenza A Viruses in Porcine Airway Epithelial Cells"

    Article Title: Differential Induction of Type I and Type III Interferons by Swine and Human Origin H1N1 Influenza A Viruses in Porcine Airway Epithelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0138704

    Replication of IAV in pAECs. (A) Primary pAECs were infected with the indicated virus at MOI of 0.01 without exogenous trypsin and the supernatants were collected at 0, 4, 8, 12, 24, 48, 72, and 96 h p.i.. Virus titers were determined by TCID 50 assay in MDCK cells in the presence of 0.5 μg/ml TPCK-trypsin. (B) Titer of IAV pH1N1 CA/09 and H1N1 IL/08 in the culture supernatant of ipAECs 72 h p.i in the presence and absence of exogenous TPCK-trypsin. Data are average of 2 independent experiments (n = 2).
    Figure Legend Snippet: Replication of IAV in pAECs. (A) Primary pAECs were infected with the indicated virus at MOI of 0.01 without exogenous trypsin and the supernatants were collected at 0, 4, 8, 12, 24, 48, 72, and 96 h p.i.. Virus titers were determined by TCID 50 assay in MDCK cells in the presence of 0.5 μg/ml TPCK-trypsin. (B) Titer of IAV pH1N1 CA/09 and H1N1 IL/08 in the culture supernatant of ipAECs 72 h p.i in the presence and absence of exogenous TPCK-trypsin. Data are average of 2 independent experiments (n = 2).

    Techniques Used: Infection

    2) Product Images from "Unique Type I Interferon Responses Determine the Functional Fate of Migratory Lung Dendritic Cells during Influenza Virus Infection"

    Article Title: Unique Type I Interferon Responses Determine the Functional Fate of Migratory Lung Dendritic Cells during Influenza Virus Infection

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1002345

    Migratory CD103 + DCs are the major cell type carrying infectious virus particles to the MLNs. A–C. Isolation of different MLN cell populations from influenza infected mice by cell sorting. A. Live gate for total MLN leukocytes shown as CD45 + DAPI - cells (gate II). B. Sorting strategy to obtain total migratory DCs (gate V), individual CD103 + DCs (gate VI), or CD11b high DCs (gate VII), and non-DC MLN cells by pooling gates i, ii, and iii during collection. All these cell populations were selected from the total CD45 + live cell gate I. C. Alternative sorting strategy starting with gate II, to obtain B220 + cells (gate B), Gr1 + cells (gate G), and pDCs (gate P2). D. Total migratory DCs (gate V) comprised of CD103 + DCs and CD11b high DCs, or MLN non-DC cells (gates I, ii, and iii pooled) sorted at 3 dpi, were layered over MDCK cells in a 1∶2 serial dilution, starting with 14,000 and 500,000 cells respectively, and were co-cultured in the presence of TPCK-trypsin for 3 days. Supernatants were assayed for infectious virus particles by hemagglutination of RBCs. D'. Supernatants from MLN-DC/MDCK co-cultures were assayed for infectious virus by plaque immunostaining on MDCK cells. E. Immunofluorescence of intracellular viral NP of sorted CD103 + DCs and CD11b high DCs from infected mice at 3 dpi. F. Individual cell populations, isolated by sorting at day 3 and 4 post-infection, as described above, were injected into 10-day old embryonated eggs, and 2 days later, allantoic fluid was assayed for infectious virus particles by hemagglutination of RBCs. Each cell type at each time point was assayed in triplicate. F’. Allantoic fluid from eggs injected with CD103 + DCs was assayed for infectious virus particles by a plaque immunostaining assay on MDCK cells.
    Figure Legend Snippet: Migratory CD103 + DCs are the major cell type carrying infectious virus particles to the MLNs. A–C. Isolation of different MLN cell populations from influenza infected mice by cell sorting. A. Live gate for total MLN leukocytes shown as CD45 + DAPI - cells (gate II). B. Sorting strategy to obtain total migratory DCs (gate V), individual CD103 + DCs (gate VI), or CD11b high DCs (gate VII), and non-DC MLN cells by pooling gates i, ii, and iii during collection. All these cell populations were selected from the total CD45 + live cell gate I. C. Alternative sorting strategy starting with gate II, to obtain B220 + cells (gate B), Gr1 + cells (gate G), and pDCs (gate P2). D. Total migratory DCs (gate V) comprised of CD103 + DCs and CD11b high DCs, or MLN non-DC cells (gates I, ii, and iii pooled) sorted at 3 dpi, were layered over MDCK cells in a 1∶2 serial dilution, starting with 14,000 and 500,000 cells respectively, and were co-cultured in the presence of TPCK-trypsin for 3 days. Supernatants were assayed for infectious virus particles by hemagglutination of RBCs. D'. Supernatants from MLN-DC/MDCK co-cultures were assayed for infectious virus by plaque immunostaining on MDCK cells. E. Immunofluorescence of intracellular viral NP of sorted CD103 + DCs and CD11b high DCs from infected mice at 3 dpi. F. Individual cell populations, isolated by sorting at day 3 and 4 post-infection, as described above, were injected into 10-day old embryonated eggs, and 2 days later, allantoic fluid was assayed for infectious virus particles by hemagglutination of RBCs. Each cell type at each time point was assayed in triplicate. F’. Allantoic fluid from eggs injected with CD103 + DCs was assayed for infectious virus particles by a plaque immunostaining assay on MDCK cells.

    Techniques Used: Isolation, Infection, Mouse Assay, FACS, Serial Dilution, Cell Culture, Immunostaining, Immunofluorescence, Injection

    Related Articles

    Infection:

    Article Title: Unique Type I Interferon Responses Determine the Functional Fate of Migratory Lung Dendritic Cells during Influenza Virus Infection
    Article Snippet: Supporting Information Migratory DCs can transfer infectious virus to uninfected cells in the absence of TPCK-trypsin. .. A. MLN-DCs (gate V, ) from PR8 or WSN infected mice (day 3) were co-cultured with MDCK cells in the presence or absence of TPCK-trypsin for 2 days. .. Virus replication was assessed by HRP-based immunostaining of MDCK cells with a chicken polyclonal antibody to influenza virus.

    Mouse Assay:

    Article Title: Unique Type I Interferon Responses Determine the Functional Fate of Migratory Lung Dendritic Cells during Influenza Virus Infection
    Article Snippet: Supporting Information Migratory DCs can transfer infectious virus to uninfected cells in the absence of TPCK-trypsin. .. A. MLN-DCs (gate V, ) from PR8 or WSN infected mice (day 3) were co-cultured with MDCK cells in the presence or absence of TPCK-trypsin for 2 days. .. Virus replication was assessed by HRP-based immunostaining of MDCK cells with a chicken polyclonal antibody to influenza virus.

    Article Title: Unique Type I Interferon Responses Determine the Functional Fate of Migratory Lung Dendritic Cells during Influenza Virus Infection
    Article Snippet: Control wells were stained with chicken normal serum followed by HRP-based immunostaining. .. B. Sorted CD103+ DCs and CD11bhigh DCs from PR8 or WSN-infected mice were co-cultured with MDCK cells in the presence or absence of TPCK-trypsin and the culture supernatants were assayed for infectious virus particles at day 2 by hemmaglutination of RBCs. (PDF) Click here for additional data file. .. Sorting strategy to isolate LN-resident CD8α+ DCs during influenza virus infection.

    Incubation:

    Article Title: Conformational Changes in the Spike Glycoprotein of Murine Coronavirus Are Induced at 37?C either by Soluble Murine CEACAM1 Receptors or by pH 8
    Article Snippet: Incubation of MHV-A59 virions at pH 6.5 and 37°C with soluble murine receptor glycoproteins CEACAM1a[1-4] (Fig. ) or the corresponding two-domain isoform smCEACAM1a[1,4] (data not shown) followed by trypsin-TPCK digestion at 4°C resulted in degradation of the S2 protein but not of the S1 protein. .. Incubation of MHV-A59 virions with smCEACAM1a[1-4] receptor glycoprotein at 4°C did not make the S2 protein susceptible to degradation by trypsin-TPCK at 4°C (Fig. ). ..

    other:

    Article Title: Conformational Changes in the Spike Glycoprotein of Murine Coronavirus Are Induced at 37?C either by Soluble Murine CEACAM1 Receptors or by pH 8
    Article Snippet: The conformational changes in S induced at 37°C by soluble receptor or pH 8.0 were apparently irreversible, since cooling the treated virions to 4°C and incubating for a prolonged time did not prevent subsequent degradation of S2 by trypsin-TPCK at 4°C.

    Article Title: Unique Type I Interferon Responses Determine the Functional Fate of Migratory Lung Dendritic Cells during Influenza Virus Infection
    Article Snippet: Supporting Information Migratory DCs can transfer infectious virus to uninfected cells in the absence of TPCK-trypsin.

    Article Title: Conformational Changes in the Spike Glycoprotein of Murine Coronavirus Are Induced at 37?C either by Soluble Murine CEACAM1 Receptors or by pH 8
    Article Snippet: As anticipated, at 37°C and pH 6.5, smCEACAM1a[1-4] induced a conformational change in the MHV-A59 spike in the SA59 R virus that made the S2 protein susceptible to degradation by trypsin-TPCK (Fig. ). smCEACAM1b[1-4] also triggered the conformational change in S2 at 37°C but did so less efficiently than did smCEACAM1a[1-4] (Fig. ).

    Article Title: Exploring protein interfaces with a general photochemical reagent
    Article Snippet: Complete enzymatic digestion of these carbamidomethylated samples with TPCK trypsin was achieved in NH4 CO3 H (0.1 M at pH 8.0), after 12–24 h at 37°C using a 2% (w/w) enzyme:substrate ratio.

    Purification:

    Article Title: The analysis of macromolecular interactions by sedimentation equilibrium
    Article Snippet: .. Purified α-chymotrypsin (Cat. No. 1475) and soybean trypsin inhibitor (STI) (Cat. No. 3570) were purchased from Worthington Biochemical Corporation. .. Approximately 21 mg of each were dissolved in 0.2M sodium chloride and 0.05M sodium phosphate (pH = 6.7) and further purified by size exclusion chromatography on a HiPrep 16/60 Sephacryl S-200 HR column (Cat. No. 17-1166-01, GE Healthcare) at 0.5 mL/min in the same buffer.

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    Worthington Biochemical tpck trypsin
    Separation by size-exclusion chromatography of tryptic peptides derived from HEWL labeled with methylene carbene. After the clean-up step described above (see Fig. ), HEWL samples were reduced, <t>carbamidomethylated,</t> and digested with <t>TPCK-trypsin</t>
    Tpck Trypsin, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tpck trypsin/product/Worthington Biochemical
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tpck trypsin - by Bioz Stars, 2021-03
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    Image Search Results


    Separation by size-exclusion chromatography of tryptic peptides derived from HEWL labeled with methylene carbene. After the clean-up step described above (see Fig. ), HEWL samples were reduced, carbamidomethylated, and digested with TPCK-trypsin

    Journal:

    Article Title: Exploring protein interfaces with a general photochemical reagent

    doi: 10.1110/ps.051960406

    Figure Lengend Snippet: Separation by size-exclusion chromatography of tryptic peptides derived from HEWL labeled with methylene carbene. After the clean-up step described above (see Fig. ), HEWL samples were reduced, carbamidomethylated, and digested with TPCK-trypsin

    Article Snippet: Complete enzymatic digestion of these carbamidomethylated samples with TPCK trypsin was achieved in NH4 CO3 H (0.1 M at pH 8.0), after 12–24 h at 37°C using a 2% (w/w) enzyme:substrate ratio.

    Techniques: Size-exclusion Chromatography, Derivative Assay, Labeling

    The S2 glycoprotein of mutant MHV-A59 virus S A59 H716D undergoes a conformational change and becomes susceptible to a unique trypsin cleavage event after incubation at 37°C with soluble murine CEACAM1a[1-4] at pH 6.5 or at pH 8.0 in the absence of receptor. Gradient purified S A59 H716D virions were incubated at 4 or 37°C for 30 min at pH 6.5 with soluble murine CEACAM1a[1-4] or without receptor proteins at pH 6.5 or 8.0. The samples were incubated at 4°C for 20 min with trypsin-TPCK at 10 μg/ml, subjected to SDS-PAGE, and immunoblotted with anti-S2 MAb 5B19.5. Untreated virions contained a novel subunit of S with an apparent molecular mass of 120 kDa. Incubation of the S A59 H716D virions with soluble murine CEACAM1a[1-4] at 37°C made the 180-kDa spike glycoprotein susceptible to cleavage by trypsin-TPCK at this novel site. To a lesser degree, incubation of virions at pH 8.0 and 37°C also increased the susceptibility of the S A59 H716D spike to trypsin cleavage.

    Journal: Journal of Virology

    Article Title: Conformational Changes in the Spike Glycoprotein of Murine Coronavirus Are Induced at 37?C either by Soluble Murine CEACAM1 Receptors or by pH 8

    doi: 10.1128/JVI.77.2.830-840.2003

    Figure Lengend Snippet: The S2 glycoprotein of mutant MHV-A59 virus S A59 H716D undergoes a conformational change and becomes susceptible to a unique trypsin cleavage event after incubation at 37°C with soluble murine CEACAM1a[1-4] at pH 6.5 or at pH 8.0 in the absence of receptor. Gradient purified S A59 H716D virions were incubated at 4 or 37°C for 30 min at pH 6.5 with soluble murine CEACAM1a[1-4] or without receptor proteins at pH 6.5 or 8.0. The samples were incubated at 4°C for 20 min with trypsin-TPCK at 10 μg/ml, subjected to SDS-PAGE, and immunoblotted with anti-S2 MAb 5B19.5. Untreated virions contained a novel subunit of S with an apparent molecular mass of 120 kDa. Incubation of the S A59 H716D virions with soluble murine CEACAM1a[1-4] at 37°C made the 180-kDa spike glycoprotein susceptible to cleavage by trypsin-TPCK at this novel site. To a lesser degree, incubation of virions at pH 8.0 and 37°C also increased the susceptibility of the S A59 H716D spike to trypsin cleavage.

    Article Snippet: As anticipated, at 37°C and pH 6.5, smCEACAM1a[1-4] induced a conformational change in the MHV-A59 spike in the SA59 R virus that made the S2 protein susceptible to degradation by trypsin-TPCK (Fig. ). smCEACAM1b[1-4] also triggered the conformational change in S2 at 37°C but did so less efficiently than did smCEACAM1a[1-4] (Fig. ).

    Techniques: Mutagenesis, Incubation, Purification, SDS Page

    Conformational changes in the S2 domain of MHV-A59 spike glycoprotein induced at 37°C by CEACAM1a at pH 6.5 or by the presence of pH 8.0 without receptor. A trypsin sensitivity assay was used to demonstrate conformational changes in the viral spike protein. Gradient-purified MHV-A59 virions were incubated at 4 or 37°C for 30 min at pH 6.5 with either soluble murine CEACAM1a[1-4] or CEACAM2[1,4] or without receptor proteins at pH 6.5 or 8.0. The samples were then incubated at 4°C for 20 min with trypsin-TPCK, subjected to SDS-PAGE, and immunoblotted with MAbs specific for the receptor binding S1 domain of the spike (MAb A1.9) or for the carboxyl-terminal S2 domain of the spike (MAb 5B93.7) which has been associated with membrane fusion.

    Journal: Journal of Virology

    Article Title: Conformational Changes in the Spike Glycoprotein of Murine Coronavirus Are Induced at 37?C either by Soluble Murine CEACAM1 Receptors or by pH 8

    doi: 10.1128/JVI.77.2.830-840.2003

    Figure Lengend Snippet: Conformational changes in the S2 domain of MHV-A59 spike glycoprotein induced at 37°C by CEACAM1a at pH 6.5 or by the presence of pH 8.0 without receptor. A trypsin sensitivity assay was used to demonstrate conformational changes in the viral spike protein. Gradient-purified MHV-A59 virions were incubated at 4 or 37°C for 30 min at pH 6.5 with either soluble murine CEACAM1a[1-4] or CEACAM2[1,4] or without receptor proteins at pH 6.5 or 8.0. The samples were then incubated at 4°C for 20 min with trypsin-TPCK, subjected to SDS-PAGE, and immunoblotted with MAbs specific for the receptor binding S1 domain of the spike (MAb A1.9) or for the carboxyl-terminal S2 domain of the spike (MAb 5B93.7) which has been associated with membrane fusion.

    Article Snippet: As anticipated, at 37°C and pH 6.5, smCEACAM1a[1-4] induced a conformational change in the MHV-A59 spike in the SA59 R virus that made the S2 protein susceptible to degradation by trypsin-TPCK (Fig. ). smCEACAM1b[1-4] also triggered the conformational change in S2 at 37°C but did so less efficiently than did smCEACAM1a[1-4] (Fig. ).

    Techniques: Sensitive Assay, Purification, Incubation, SDS Page, Binding Assay

    Migratory CD103 + DCs are the major cell type carrying infectious virus particles to the MLNs. A–C. Isolation of different MLN cell populations from influenza infected mice by cell sorting. A. Live gate for total MLN leukocytes shown as CD45 + DAPI - cells (gate II). B. Sorting strategy to obtain total migratory DCs (gate V), individual CD103 + DCs (gate VI), or CD11b high DCs (gate VII), and non-DC MLN cells by pooling gates i, ii, and iii during collection. All these cell populations were selected from the total CD45 + live cell gate I. C. Alternative sorting strategy starting with gate II, to obtain B220 + cells (gate B), Gr1 + cells (gate G), and pDCs (gate P2). D. Total migratory DCs (gate V) comprised of CD103 + DCs and CD11b high DCs, or MLN non-DC cells (gates I, ii, and iii pooled) sorted at 3 dpi, were layered over MDCK cells in a 1∶2 serial dilution, starting with 14,000 and 500,000 cells respectively, and were co-cultured in the presence of TPCK-trypsin for 3 days. Supernatants were assayed for infectious virus particles by hemagglutination of RBCs. D'. Supernatants from MLN-DC/MDCK co-cultures were assayed for infectious virus by plaque immunostaining on MDCK cells. E. Immunofluorescence of intracellular viral NP of sorted CD103 + DCs and CD11b high DCs from infected mice at 3 dpi. F. Individual cell populations, isolated by sorting at day 3 and 4 post-infection, as described above, were injected into 10-day old embryonated eggs, and 2 days later, allantoic fluid was assayed for infectious virus particles by hemagglutination of RBCs. Each cell type at each time point was assayed in triplicate. F’. Allantoic fluid from eggs injected with CD103 + DCs was assayed for infectious virus particles by a plaque immunostaining assay on MDCK cells.

    Journal: PLoS Pathogens

    Article Title: Unique Type I Interferon Responses Determine the Functional Fate of Migratory Lung Dendritic Cells during Influenza Virus Infection

    doi: 10.1371/journal.ppat.1002345

    Figure Lengend Snippet: Migratory CD103 + DCs are the major cell type carrying infectious virus particles to the MLNs. A–C. Isolation of different MLN cell populations from influenza infected mice by cell sorting. A. Live gate for total MLN leukocytes shown as CD45 + DAPI - cells (gate II). B. Sorting strategy to obtain total migratory DCs (gate V), individual CD103 + DCs (gate VI), or CD11b high DCs (gate VII), and non-DC MLN cells by pooling gates i, ii, and iii during collection. All these cell populations were selected from the total CD45 + live cell gate I. C. Alternative sorting strategy starting with gate II, to obtain B220 + cells (gate B), Gr1 + cells (gate G), and pDCs (gate P2). D. Total migratory DCs (gate V) comprised of CD103 + DCs and CD11b high DCs, or MLN non-DC cells (gates I, ii, and iii pooled) sorted at 3 dpi, were layered over MDCK cells in a 1∶2 serial dilution, starting with 14,000 and 500,000 cells respectively, and were co-cultured in the presence of TPCK-trypsin for 3 days. Supernatants were assayed for infectious virus particles by hemagglutination of RBCs. D'. Supernatants from MLN-DC/MDCK co-cultures were assayed for infectious virus by plaque immunostaining on MDCK cells. E. Immunofluorescence of intracellular viral NP of sorted CD103 + DCs and CD11b high DCs from infected mice at 3 dpi. F. Individual cell populations, isolated by sorting at day 3 and 4 post-infection, as described above, were injected into 10-day old embryonated eggs, and 2 days later, allantoic fluid was assayed for infectious virus particles by hemagglutination of RBCs. Each cell type at each time point was assayed in triplicate. F’. Allantoic fluid from eggs injected with CD103 + DCs was assayed for infectious virus particles by a plaque immunostaining assay on MDCK cells.

    Article Snippet: Supporting Information Migratory DCs can transfer infectious virus to uninfected cells in the absence of TPCK-trypsin.

    Techniques: Isolation, Infection, Mouse Assay, FACS, Serial Dilution, Cell Culture, Immunostaining, Immunofluorescence, Injection