trypsin tpck  (Worthington Biochemical)


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    Name:
    Trypsin TPCK Treated
    Description:
    A chromatographically purified diafiltered lyophilized powder that has been treated with L tosylamido 2 phenyl ethyl chloromethyl ketone TPCK to inhibit contaminating chymotryptic activity Kostka and Carpenter J Biol Chem 239 1799 1964
    Catalog Number:
    LS003740
    Price:
    86
    Source:
    Bovine Pancreas
    Size:
    100 mg
    Cas Number:
    9002.07.7
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    Structured Review

    Worthington Biochemical trypsin tpck
    The R377W and F488L mutations do not affect protein stability. (A). Thermal denaturation study of the WT and R377W and F488L mutant C-termini. Shown are thermal denaturation curves for the WT and mutant C-termini obtained at 220 nm from 20°C to 80°C (GST, black; WT, blue; R377W, green; and F488L, red). Denaturation curves are in units of molar ellipticity vs. temperature. The transition temperature was at ∼ 59°C for GST and was at ∼ 68°C for the C-terminal fusion proteins. The unfolding temperature at the R377W and F488L mutants was not different from that in the WT. (B). Limited tryptic digestion of the WT and R377W and F488L mutant CNGA3. The membranes prepared from cells that had been <t>transfected</t> with cDNAs encoding the WT and R377W and F488L mutants were incubated with <t>trypsin-TPCK</t> (30 μg/ml) at 30°C for 2, 5, and 10 min. The digested products were resolved on 10% SDS-PAGE, followed by Western blotting using the polyclonal anti-CNGA3 antibody. Shown are representative images showing limited proteolysis of the WT and mutant CNGA3 subunits. The cleavage rates of the mutants to trypsin were about the same as the WT.
    A chromatographically purified diafiltered lyophilized powder that has been treated with L tosylamido 2 phenyl ethyl chloromethyl ketone TPCK to inhibit contaminating chymotryptic activity Kostka and Carpenter J Biol Chem 239 1799 1964
    https://www.bioz.com/result/trypsin tpck/product/Worthington Biochemical
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trypsin tpck - by Bioz Stars, 2021-07
    97/100 stars

    Images

    1) Product Images from "The Disease-Causing Mutations in the Carboxyl-Terminus of Cone Cyclic Nucleotide-Gated Channel CNGA3 Subunit Alter the Local Secondary Structure and Interfere with the Channel Active Conformational Change"

    Article Title: The Disease-Causing Mutations in the Carboxyl-Terminus of Cone Cyclic Nucleotide-Gated Channel CNGA3 Subunit Alter the Local Secondary Structure and Interfere with the Channel Active Conformational Change

    Journal: Biochemistry

    doi: 10.1021/bi901960u

    The R377W and F488L mutations do not affect protein stability. (A). Thermal denaturation study of the WT and R377W and F488L mutant C-termini. Shown are thermal denaturation curves for the WT and mutant C-termini obtained at 220 nm from 20°C to 80°C (GST, black; WT, blue; R377W, green; and F488L, red). Denaturation curves are in units of molar ellipticity vs. temperature. The transition temperature was at ∼ 59°C for GST and was at ∼ 68°C for the C-terminal fusion proteins. The unfolding temperature at the R377W and F488L mutants was not different from that in the WT. (B). Limited tryptic digestion of the WT and R377W and F488L mutant CNGA3. The membranes prepared from cells that had been transfected with cDNAs encoding the WT and R377W and F488L mutants were incubated with trypsin-TPCK (30 μg/ml) at 30°C for 2, 5, and 10 min. The digested products were resolved on 10% SDS-PAGE, followed by Western blotting using the polyclonal anti-CNGA3 antibody. Shown are representative images showing limited proteolysis of the WT and mutant CNGA3 subunits. The cleavage rates of the mutants to trypsin were about the same as the WT.
    Figure Legend Snippet: The R377W and F488L mutations do not affect protein stability. (A). Thermal denaturation study of the WT and R377W and F488L mutant C-termini. Shown are thermal denaturation curves for the WT and mutant C-termini obtained at 220 nm from 20°C to 80°C (GST, black; WT, blue; R377W, green; and F488L, red). Denaturation curves are in units of molar ellipticity vs. temperature. The transition temperature was at ∼ 59°C for GST and was at ∼ 68°C for the C-terminal fusion proteins. The unfolding temperature at the R377W and F488L mutants was not different from that in the WT. (B). Limited tryptic digestion of the WT and R377W and F488L mutant CNGA3. The membranes prepared from cells that had been transfected with cDNAs encoding the WT and R377W and F488L mutants were incubated with trypsin-TPCK (30 μg/ml) at 30°C for 2, 5, and 10 min. The digested products were resolved on 10% SDS-PAGE, followed by Western blotting using the polyclonal anti-CNGA3 antibody. Shown are representative images showing limited proteolysis of the WT and mutant CNGA3 subunits. The cleavage rates of the mutants to trypsin were about the same as the WT.

    Techniques Used: Mutagenesis, Transfection, Incubation, SDS Page, Western Blot

    Related Articles

    Purification:

    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
    Article Snippet: The swine influenza viruses; A/Sw/Illinois/2008 (H1N1 IL/08), A/Sw/Iowa/2004 (H1N1 IA/04), and A (H3N2) variant virus (H3N2v), and the human pandemic influenza A/California/04/2009 (pH1N1 CA/09) were obtained from the University of Minnesota Veterinary Diagnostic Laboratory (St Paul, MN). .. Viruses were propagated in MDCK cells in DMEM containing 0.5 μg/ml TPCK-trypsin (Worthington Biochemical Corporation, Lakewood, NJ) and purified from the clarified cell culture supernatants by ultracentrifugation through a 30% (w/v) sucrose cushion and stored in aliquots at -80o C. Culture supernatant from un-infected MDCK cells were processed similarly to use for mock infection. .. Isolation of primary pAECs Primary pAECs were isolated from three healthy neonatal pigs as described previously [ ] with slight modifications.

    Cell Culture:

    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
    Article Snippet: The swine influenza viruses; A/Sw/Illinois/2008 (H1N1 IL/08), A/Sw/Iowa/2004 (H1N1 IA/04), and A (H3N2) variant virus (H3N2v), and the human pandemic influenza A/California/04/2009 (pH1N1 CA/09) were obtained from the University of Minnesota Veterinary Diagnostic Laboratory (St Paul, MN). .. Viruses were propagated in MDCK cells in DMEM containing 0.5 μg/ml TPCK-trypsin (Worthington Biochemical Corporation, Lakewood, NJ) and purified from the clarified cell culture supernatants by ultracentrifugation through a 30% (w/v) sucrose cushion and stored in aliquots at -80o C. Culture supernatant from un-infected MDCK cells were processed similarly to use for mock infection. .. Isolation of primary pAECs Primary pAECs were isolated from three healthy neonatal pigs as described previously [ ] with slight modifications.

    Infection:

    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
    Article Snippet: The swine influenza viruses; A/Sw/Illinois/2008 (H1N1 IL/08), A/Sw/Iowa/2004 (H1N1 IA/04), and A (H3N2) variant virus (H3N2v), and the human pandemic influenza A/California/04/2009 (pH1N1 CA/09) were obtained from the University of Minnesota Veterinary Diagnostic Laboratory (St Paul, MN). .. Viruses were propagated in MDCK cells in DMEM containing 0.5 μg/ml TPCK-trypsin (Worthington Biochemical Corporation, Lakewood, NJ) and purified from the clarified cell culture supernatants by ultracentrifugation through a 30% (w/v) sucrose cushion and stored in aliquots at -80o C. Culture supernatant from un-infected MDCK cells were processed similarly to use for mock infection. .. Isolation of primary pAECs Primary pAECs were isolated from three healthy neonatal pigs as described previously [ ] with slight modifications.

    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.

    other:

    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.

    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.

    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. ). ..

    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: Surprisingly, a trypsin sensitivity assay of the SA59 H716D mutant virus (Fig. ) showed that after incubation at pH 6.5 and 4 or 37°C, a minor fraction of the spike glycoprotein of the SA59 H716D virus could be proteolytically cleaved by trypsin at 4°C to yield a novel 120-kDa subunit that was detected by MAbs to both S1 and S2. .. When SA59 H716D virions were incubated at 37°C and pH 6.5 with smCEACAM1a[1-4] or at pH 8.0 without receptor, the S glycoprotein became highly susceptible to cleavage with trypsin-TPCK at 4°C to yield the 120-kDa glycoprotein, without further degradation of S2 as seen with wild-type MHV-A59 and SA59 R, (Fig. and , respectively). ..

    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.

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  • 97
    Worthington Biochemical trypsin tpck
    The R377W and F488L mutations do not affect protein stability. (A). Thermal denaturation study of the WT and R377W and F488L mutant C-termini. Shown are thermal denaturation curves for the WT and mutant C-termini obtained at 220 nm from 20°C to 80°C (GST, black; WT, blue; R377W, green; and F488L, red). Denaturation curves are in units of molar ellipticity vs. temperature. The transition temperature was at ∼ 59°C for GST and was at ∼ 68°C for the C-terminal fusion proteins. The unfolding temperature at the R377W and F488L mutants was not different from that in the WT. (B). Limited tryptic digestion of the WT and R377W and F488L mutant CNGA3. The membranes prepared from cells that had been <t>transfected</t> with cDNAs encoding the WT and R377W and F488L mutants were incubated with <t>trypsin-TPCK</t> (30 μg/ml) at 30°C for 2, 5, and 10 min. The digested products were resolved on 10% SDS-PAGE, followed by Western blotting using the polyclonal anti-CNGA3 antibody. Shown are representative images showing limited proteolysis of the WT and mutant CNGA3 subunits. The cleavage rates of the mutants to trypsin were about the same as the WT.
    Trypsin Tpck, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trypsin tpck/product/Worthington Biochemical
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trypsin tpck - by Bioz Stars, 2021-07
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    99
    Worthington Biochemical dnase i
    Human and mouse necroptotic neutrophils release NETs that are sensitive to <t>DNase</t> I and inhibited by Nec-1s. ( A and B . FasL, Fas ligand; BF, bright field. ( C ) Flow cytometric analysis of membrane permeability and PS exposure assessed by propidium iodide and annexin V staining of neutrophils stimulated for 12 hours as indicated. Data are representative of five independent experiments. G-CSF, granulocyte colony-stimulating factor. ( D to H ) Flow cytometric analysis of cellular viability and NET formation assessed by extracellular DNA and H3Cit staining of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. Deoxyribonuclease I (DNase I) pretreatment eliminated the appearance of H3Cit + PicoGreen + NET-producing cells (E), without effecting PicoGreen − viable cells (F). Representative dot plots (D) were quantified (E and F), and data are means ± SEM of four independent experiments. ( G and H ) Flow cytometric analysis of cellular viability and NET formation of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. NET formation (G) and viability (H) are means ± SEM of three independent experiments. * P
    Dnase I, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Worthington Biochemical mg ml 1 collagenase
    Human and mouse necroptotic neutrophils release NETs that are sensitive to <t>DNase</t> I and inhibited by Nec-1s. ( A and B . FasL, Fas ligand; BF, bright field. ( C ) Flow cytometric analysis of membrane permeability and PS exposure assessed by propidium iodide and annexin V staining of neutrophils stimulated for 12 hours as indicated. Data are representative of five independent experiments. G-CSF, granulocyte colony-stimulating factor. ( D to H ) Flow cytometric analysis of cellular viability and NET formation assessed by extracellular DNA and H3Cit staining of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. Deoxyribonuclease I (DNase I) pretreatment eliminated the appearance of H3Cit + PicoGreen + NET-producing cells (E), without effecting PicoGreen − viable cells (F). Representative dot plots (D) were quantified (E and F), and data are means ± SEM of four independent experiments. ( G and H ) Flow cytometric analysis of cellular viability and NET formation of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. NET formation (G) and viability (H) are means ± SEM of three independent experiments. * P
    Mg Ml 1 Collagenase, supplied by Worthington Biochemical, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mg ml 1 collagenase/product/Worthington Biochemical
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    Image Search Results


    The R377W and F488L mutations do not affect protein stability. (A). Thermal denaturation study of the WT and R377W and F488L mutant C-termini. Shown are thermal denaturation curves for the WT and mutant C-termini obtained at 220 nm from 20°C to 80°C (GST, black; WT, blue; R377W, green; and F488L, red). Denaturation curves are in units of molar ellipticity vs. temperature. The transition temperature was at ∼ 59°C for GST and was at ∼ 68°C for the C-terminal fusion proteins. The unfolding temperature at the R377W and F488L mutants was not different from that in the WT. (B). Limited tryptic digestion of the WT and R377W and F488L mutant CNGA3. The membranes prepared from cells that had been transfected with cDNAs encoding the WT and R377W and F488L mutants were incubated with trypsin-TPCK (30 μg/ml) at 30°C for 2, 5, and 10 min. The digested products were resolved on 10% SDS-PAGE, followed by Western blotting using the polyclonal anti-CNGA3 antibody. Shown are representative images showing limited proteolysis of the WT and mutant CNGA3 subunits. The cleavage rates of the mutants to trypsin were about the same as the WT.

    Journal: Biochemistry

    Article Title: The Disease-Causing Mutations in the Carboxyl-Terminus of Cone Cyclic Nucleotide-Gated Channel CNGA3 Subunit Alter the Local Secondary Structure and Interfere with the Channel Active Conformational Change

    doi: 10.1021/bi901960u

    Figure Lengend Snippet: The R377W and F488L mutations do not affect protein stability. (A). Thermal denaturation study of the WT and R377W and F488L mutant C-termini. Shown are thermal denaturation curves for the WT and mutant C-termini obtained at 220 nm from 20°C to 80°C (GST, black; WT, blue; R377W, green; and F488L, red). Denaturation curves are in units of molar ellipticity vs. temperature. The transition temperature was at ∼ 59°C for GST and was at ∼ 68°C for the C-terminal fusion proteins. The unfolding temperature at the R377W and F488L mutants was not different from that in the WT. (B). Limited tryptic digestion of the WT and R377W and F488L mutant CNGA3. The membranes prepared from cells that had been transfected with cDNAs encoding the WT and R377W and F488L mutants were incubated with trypsin-TPCK (30 μg/ml) at 30°C for 2, 5, and 10 min. The digested products were resolved on 10% SDS-PAGE, followed by Western blotting using the polyclonal anti-CNGA3 antibody. Shown are representative images showing limited proteolysis of the WT and mutant CNGA3 subunits. The cleavage rates of the mutants to trypsin were about the same as the WT.

    Article Snippet: The membranes prepared from cells that had been transfected with cDNAs for the WT and mutants were incubated with trypsin-TPCK (30 μg/ml) at 30°C for 2, 5, and 10 min.

    Techniques: Mutagenesis, Transfection, Incubation, SDS Page, Western Blot

    Human and mouse necroptotic neutrophils release NETs that are sensitive to DNase I and inhibited by Nec-1s. ( A and B . FasL, Fas ligand; BF, bright field. ( C ) Flow cytometric analysis of membrane permeability and PS exposure assessed by propidium iodide and annexin V staining of neutrophils stimulated for 12 hours as indicated. Data are representative of five independent experiments. G-CSF, granulocyte colony-stimulating factor. ( D to H ) Flow cytometric analysis of cellular viability and NET formation assessed by extracellular DNA and H3Cit staining of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. Deoxyribonuclease I (DNase I) pretreatment eliminated the appearance of H3Cit + PicoGreen + NET-producing cells (E), without effecting PicoGreen − viable cells (F). Representative dot plots (D) were quantified (E and F), and data are means ± SEM of four independent experiments. ( G and H ) Flow cytometric analysis of cellular viability and NET formation of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. NET formation (G) and viability (H) are means ± SEM of three independent experiments. * P

    Journal: Science signaling

    Article Title: The pseudokinase MLKL activates PAD4-dependent NET formation in necroptotic neutrophils

    doi: 10.1126/scisignal.aao1716

    Figure Lengend Snippet: Human and mouse necroptotic neutrophils release NETs that are sensitive to DNase I and inhibited by Nec-1s. ( A and B . FasL, Fas ligand; BF, bright field. ( C ) Flow cytometric analysis of membrane permeability and PS exposure assessed by propidium iodide and annexin V staining of neutrophils stimulated for 12 hours as indicated. Data are representative of five independent experiments. G-CSF, granulocyte colony-stimulating factor. ( D to H ) Flow cytometric analysis of cellular viability and NET formation assessed by extracellular DNA and H3Cit staining of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. Deoxyribonuclease I (DNase I) pretreatment eliminated the appearance of H3Cit + PicoGreen + NET-producing cells (E), without effecting PicoGreen − viable cells (F). Representative dot plots (D) were quantified (E and F), and data are means ± SEM of four independent experiments. ( G and H ) Flow cytometric analysis of cellular viability and NET formation of IFN-γ–primed mouse bone marrow neutrophils treated as indicated. NET formation (G) and viability (H) are means ± SEM of three independent experiments. * P

    Article Snippet: MRSA [multiplicity of infection (MOI), 1] was added to cells for a further 30 min in the presence or absence of DNase I (10 U/ml).

    Techniques: Flow Cytometry, Permeability, Staining

    DNA binding activity and footprinting analysis. ( A ) EMSA using radioactively labeled single-stranded pSLA2 telomeric DNA. (Lane a ) Probe + BSA as control. (Lanes b,c,d ) TpgL protein with 5-fold, 20-fold, and 100-fold excesses of single-stranded BKKO5 DNA. (Lanes e,g,f ) Same as lanes b, c , and d except Tap L protein was used. ( B ) EMSA using radioactively labeled single-stranded pSLA2 telomeric DNA in the presence of unlabeled cold probe as competitor. (Lane a ) Probe + BSA as control. (Lanes b,c,d ) TpgL protein with 5-fold, 20-fold, and 100-fold excesses of single-stranded cold probe. (Lanes e,g,f ) Same as lanes b, c , and d except Tap L protein was used. ( C ) EMSA using radioactively labeled double-stranded pSLA2 telomeric DNA. (Lane a ) Probe + BSA as control. (Lanes b,c,d ) TpgL protein with 5-fold, 20-fold, and 100-fold excesses of circular chromosomal DNA of BKKO5. (Lane e ) Tap L protein alone. ( D ) DNase I footprinting analysis using single-stranded pSLA2 telomeric DNA and Tap L protein was carried out as described in Materials and Methods. (Lanes 1,2 ) Single-stranded telomeric DNA was incubated with 0 and 2 μg of Tap L protein, respectively. (Lanes 3 – 6 ) DNA sequencing ladder reactions with termination mix of ddG, ddA, ddT, and ddC. Brackets at the right show two regions of the protected DNA sequences.

    Journal: Genes & Development

    Article Title: Recruitment of terminal protein to the ends of Streptomyces linear plasmids and chromosomes by a novel telomere-binding protein essential for linear DNA replication

    doi: 10.1101/gad.1060303

    Figure Lengend Snippet: DNA binding activity and footprinting analysis. ( A ) EMSA using radioactively labeled single-stranded pSLA2 telomeric DNA. (Lane a ) Probe + BSA as control. (Lanes b,c,d ) TpgL protein with 5-fold, 20-fold, and 100-fold excesses of single-stranded BKKO5 DNA. (Lanes e,g,f ) Same as lanes b, c , and d except Tap L protein was used. ( B ) EMSA using radioactively labeled single-stranded pSLA2 telomeric DNA in the presence of unlabeled cold probe as competitor. (Lane a ) Probe + BSA as control. (Lanes b,c,d ) TpgL protein with 5-fold, 20-fold, and 100-fold excesses of single-stranded cold probe. (Lanes e,g,f ) Same as lanes b, c , and d except Tap L protein was used. ( C ) EMSA using radioactively labeled double-stranded pSLA2 telomeric DNA. (Lane a ) Probe + BSA as control. (Lanes b,c,d ) TpgL protein with 5-fold, 20-fold, and 100-fold excesses of circular chromosomal DNA of BKKO5. (Lane e ) Tap L protein alone. ( D ) DNase I footprinting analysis using single-stranded pSLA2 telomeric DNA and Tap L protein was carried out as described in Materials and Methods. (Lanes 1,2 ) Single-stranded telomeric DNA was incubated with 0 and 2 μg of Tap L protein, respectively. (Lanes 3 – 6 ) DNA sequencing ladder reactions with termination mix of ddG, ddA, ddT, and ddC. Brackets at the right show two regions of the protected DNA sequences.

    Article Snippet: CaCl2 and MgCl2 were added to final concentrations of 2.5 and 5 mM, respectively, and 1 μL DNase I (10 μg/mL) was added.

    Techniques: Binding Assay, Activity Assay, Footprinting, Labeling, Incubation, DNA Sequencing