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Bio-Rad proteinase k
Characterization of RiCF. (A)  Representative radiogram showing RNase dose dependent chromosomal fragmentation in AB1157. Plugs were made with 0, 2, 10, 25, 50 or 100 μg RNase and lysed and electrophoresed under standard conditions. CZ, compression zone.  (B)  Quantification showing increase in chromosomal fragmentation in RNase dose-dependent manner. Data points are means of six independent assays ± SEM.  (C)  RNase-effect is not seen in the pre-lyzed cells. Plugs from AB1157 culture were made in the presence of proteinase K, but without any RNase. After overnight lysis and extensive washing, the plugs were incubated with 0, 2, 20 and 100 μg RNase or 100 U of EcoRI for 15 H at 37°C before PFGE.  (D)  Quantification of chromosomal fragmentation showing extreme sensitivity of chromosomes to EcoRI, but not RNase, when plugs were treated with the enzymes after lysis of cells. The experiment is done twice and a representative result is presented.  (E)  A representative radiogram showing kinetics of RiCF. Multiple plugs were made in the presence of RNase (50 μg/plug) and incubated at 62°C for 10, 30, 60, 180 or 900 minutes with lysis buffer in individual tubes. At the indicated times, one tube was removed, lysis buffer was replaced with ice-cold TE, and plugs were stored at 4°C until all plugs were ready for electrophoresis.  (F)  Quantification of kinetics of chromosomal fragmentation when plugs were made in the presence of RNase and lysed for 1, 5, 10, 30, 60, 180 or 900 minutes. Data points are means of three independent assays ± SEM. Arrow shows the value of fragmentation after 10 min lysis.
Proteinase K, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Degradation of RNA during lysis of Escherichia coli cells in agarose plugs breaks the chromosome"

Article Title: Degradation of RNA during lysis of Escherichia coli cells in agarose plugs breaks the chromosome

Journal: PLoS ONE

doi: 10.1371/journal.pone.0190177

Characterization of RiCF. (A)  Representative radiogram showing RNase dose dependent chromosomal fragmentation in AB1157. Plugs were made with 0, 2, 10, 25, 50 or 100 μg RNase and lysed and electrophoresed under standard conditions. CZ, compression zone.  (B)  Quantification showing increase in chromosomal fragmentation in RNase dose-dependent manner. Data points are means of six independent assays ± SEM.  (C)  RNase-effect is not seen in the pre-lyzed cells. Plugs from AB1157 culture were made in the presence of proteinase K, but without any RNase. After overnight lysis and extensive washing, the plugs were incubated with 0, 2, 20 and 100 μg RNase or 100 U of EcoRI for 15 H at 37°C before PFGE.  (D)  Quantification of chromosomal fragmentation showing extreme sensitivity of chromosomes to EcoRI, but not RNase, when plugs were treated with the enzymes after lysis of cells. The experiment is done twice and a representative result is presented.  (E)  A representative radiogram showing kinetics of RiCF. Multiple plugs were made in the presence of RNase (50 μg/plug) and incubated at 62°C for 10, 30, 60, 180 or 900 minutes with lysis buffer in individual tubes. At the indicated times, one tube was removed, lysis buffer was replaced with ice-cold TE, and plugs were stored at 4°C until all plugs were ready for electrophoresis.  (F)  Quantification of kinetics of chromosomal fragmentation when plugs were made in the presence of RNase and lysed for 1, 5, 10, 30, 60, 180 or 900 minutes. Data points are means of three independent assays ± SEM. Arrow shows the value of fragmentation after 10 min lysis.
Figure Legend Snippet: Characterization of RiCF. (A) Representative radiogram showing RNase dose dependent chromosomal fragmentation in AB1157. Plugs were made with 0, 2, 10, 25, 50 or 100 μg RNase and lysed and electrophoresed under standard conditions. CZ, compression zone. (B) Quantification showing increase in chromosomal fragmentation in RNase dose-dependent manner. Data points are means of six independent assays ± SEM. (C) RNase-effect is not seen in the pre-lyzed cells. Plugs from AB1157 culture were made in the presence of proteinase K, but without any RNase. After overnight lysis and extensive washing, the plugs were incubated with 0, 2, 20 and 100 μg RNase or 100 U of EcoRI for 15 H at 37°C before PFGE. (D) Quantification of chromosomal fragmentation showing extreme sensitivity of chromosomes to EcoRI, but not RNase, when plugs were treated with the enzymes after lysis of cells. The experiment is done twice and a representative result is presented. (E) A representative radiogram showing kinetics of RiCF. Multiple plugs were made in the presence of RNase (50 μg/plug) and incubated at 62°C for 10, 30, 60, 180 or 900 minutes with lysis buffer in individual tubes. At the indicated times, one tube was removed, lysis buffer was replaced with ice-cold TE, and plugs were stored at 4°C until all plugs were ready for electrophoresis. (F) Quantification of kinetics of chromosomal fragmentation when plugs were made in the presence of RNase and lysed for 1, 5, 10, 30, 60, 180 or 900 minutes. Data points are means of three independent assays ± SEM. Arrow shows the value of fragmentation after 10 min lysis.

Techniques Used: Lysis, Incubation, Electrophoresis

Genetics of RiCF. (A)  Quantitative determination of RiCF in Δ hns  mutant. AB1157 and SRK254 were grown at 37°C to the same final OD, and plugs were made in the absence of proteinase K, but with or without RNase (50 μg/plug). After overnight incubation in the lysis buffer at 62°C, the plugs were electrophoresed under standard conditions. Data points are means of 6–10 independent assays± SEM.  (B)  Radiogram of a representative pulsed field gel from which data in (A) are calculated.  (C)  Comparison of RiCF in Δ hupA  Δ hupB  and Δ ihfA  Δ ihfB  double mutants. Experiment was done as described in (A), and values presented are means of 6–13 independent assays ± SEM.  (D)  Radiogram of a representative pulsed field gel from which data in (C) are calculated.  (E)  Effect of  hns  deletion on RiCF of Δ ihfA  Δ ihfB  mutant. Values presented are means of 7 independent assays ± SEM.
Figure Legend Snippet: Genetics of RiCF. (A) Quantitative determination of RiCF in Δ hns mutant. AB1157 and SRK254 were grown at 37°C to the same final OD, and plugs were made in the absence of proteinase K, but with or without RNase (50 μg/plug). After overnight incubation in the lysis buffer at 62°C, the plugs were electrophoresed under standard conditions. Data points are means of 6–10 independent assays± SEM. (B) Radiogram of a representative pulsed field gel from which data in (A) are calculated. (C) Comparison of RiCF in Δ hupA Δ hupB and Δ ihfA Δ ihfB double mutants. Experiment was done as described in (A), and values presented are means of 6–13 independent assays ± SEM. (D) Radiogram of a representative pulsed field gel from which data in (C) are calculated. (E) Effect of hns deletion on RiCF of Δ ihfA Δ ihfB mutant. Values presented are means of 7 independent assays ± SEM.

Techniques Used: Mutagenesis, Incubation, Lysis, Pulsed-Field Gel

Non-coding RNA and HU stabilize nucleoids. (A)  Comparison of spontaneous and RNase-induced fragmentation in Δ nc1  Δ nc5  Δ ihfA  Δ ihfB , Δ nc1  Δ nc5  Δ hupA  Δ hupB  and Δ nc1  Δ nc5  Δ hns  mutants. AB1157, SRK254-12, SRK254-15 and SRK254-18 were grown at 37°C, and plugs were made in absence of proteinase K, both with or without RNase (50 μg/plug), and lysed under standard conditions. Data points are means of at least three independent assays± SEM.  (B)  Radiogram of a representative gel from which data in (A) are generated.
Figure Legend Snippet: Non-coding RNA and HU stabilize nucleoids. (A) Comparison of spontaneous and RNase-induced fragmentation in Δ nc1 Δ nc5 Δ ihfA Δ ihfB , Δ nc1 Δ nc5 Δ hupA Δ hupB and Δ nc1 Δ nc5 Δ hns mutants. AB1157, SRK254-12, SRK254-15 and SRK254-18 were grown at 37°C, and plugs were made in absence of proteinase K, both with or without RNase (50 μg/plug), and lysed under standard conditions. Data points are means of at least three independent assays± SEM. (B) Radiogram of a representative gel from which data in (A) are generated.

Techniques Used: Generated

Endonuclease-I is critical for RiCF but not for spontaneous fragmentation. (A)  Comparison of spontaneous fragmentation and RiCF in AB1157, AB1157 Δ nc1  Δ nc5  Δ hupA  Δ hupB  and AB1157 Δ nc1  Δ nc5  Δ hupA  Δ hupB  Δ endA  mutants. All strains were grown at 37°C to the final OD of 0.6, and plugs were made in the absence of proteinase K, both with or without RNase (50 μg/plug). Data points are means of at least three independent assays ± SEM.  (B)  Radiogram of a representative gel from which data in (A) are generated.
Figure Legend Snippet: Endonuclease-I is critical for RiCF but not for spontaneous fragmentation. (A) Comparison of spontaneous fragmentation and RiCF in AB1157, AB1157 Δ nc1 Δ nc5 Δ hupA Δ hupB and AB1157 Δ nc1 Δ nc5 Δ hupA Δ hupB Δ endA mutants. All strains were grown at 37°C to the final OD of 0.6, and plugs were made in the absence of proteinase K, both with or without RNase (50 μg/plug). Data points are means of at least three independent assays ± SEM. (B) Radiogram of a representative gel from which data in (A) are generated.

Techniques Used: Generated

RNA degradation causes chromosomal fragmentation. (A) Schematics of a hypothetical scenario when RNA makes the central core of nucleoids, and its degradation results in collapse of the nucleoid structure, causing chromosomal fragmentation. (B) Radiogram of a pulsed field gel showing chromosomal fragmentation in AB1157 when cells were embedded in agarose plugs in the presence and absence of proteinase K (25 μg/plug) and/or RNase (50 μg/plug) and lysed overnight at 62°C. (C) Radiogram showing DNase I sensitivity of the signal entering the gel. Plugs were lysed at 62°C, washed extensively to remove traces of lysis buffer and then treated with DNase I at 37°C before PFGE. (D) A representative gel showing that RNA degradation by different enzymes causes chromosomal fragmentation. Plugs were made in the absence of proteinase K in 1x restriction enzyme buffer (NEBuffer 3 for RNase A, XRN-1 and RNAse I f and NEBuffer 4 for Exo T). The concentrations of the enzymes used were, RNase, 50 μg/plug; XRN-1, 5 U/plug; RNAse I f , 100 U/plug and Exo T, 20 U/plug. (E) Quantification of the chromosomal fragmentation when plugs were made in the presence of various RNA degrading enzymes. The values presented are means of four independent assays ± SEM. CZ, compression zone.
Figure Legend Snippet: RNA degradation causes chromosomal fragmentation. (A) Schematics of a hypothetical scenario when RNA makes the central core of nucleoids, and its degradation results in collapse of the nucleoid structure, causing chromosomal fragmentation. (B) Radiogram of a pulsed field gel showing chromosomal fragmentation in AB1157 when cells were embedded in agarose plugs in the presence and absence of proteinase K (25 μg/plug) and/or RNase (50 μg/plug) and lysed overnight at 62°C. (C) Radiogram showing DNase I sensitivity of the signal entering the gel. Plugs were lysed at 62°C, washed extensively to remove traces of lysis buffer and then treated with DNase I at 37°C before PFGE. (D) A representative gel showing that RNA degradation by different enzymes causes chromosomal fragmentation. Plugs were made in the absence of proteinase K in 1x restriction enzyme buffer (NEBuffer 3 for RNase A, XRN-1 and RNAse I f and NEBuffer 4 for Exo T). The concentrations of the enzymes used were, RNase, 50 μg/plug; XRN-1, 5 U/plug; RNAse I f , 100 U/plug and Exo T, 20 U/plug. (E) Quantification of the chromosomal fragmentation when plugs were made in the presence of various RNA degrading enzymes. The values presented are means of four independent assays ± SEM. CZ, compression zone.

Techniques Used: Pulsed-Field Gel, Lysis

2) Product Images from "The majority of in vitro macrophage activation exhibited by extracts of some immune enhancing botanicals is due to bacterial lipoproteins and lipopolysaccharides"

Article Title: The majority of in vitro macrophage activation exhibited by extracts of some immune enhancing botanicals is due to bacterial lipoproteins and lipopolysaccharides

Journal: International immunopharmacology

doi: 10.1016/j.intimp.2008.03.007

Treatment with proteinase K reduces the size of the active compounds in the melanin fraction of botanicals. The melanin fraction of the botanicals indicated was dissolved in 4% SDS and treated with proteinase K (black bars) or left untreated (white bars).
Figure Legend Snippet: Treatment with proteinase K reduces the size of the active compounds in the melanin fraction of botanicals. The melanin fraction of the botanicals indicated was dissolved in 4% SDS and treated with proteinase K (black bars) or left untreated (white bars).

Techniques Used:

Identification within Echinacea purpurea root of 2,3-dihydroxypropyl cysteine, a structural component unique to prokaryotic lipoproteins. Lipoproteins were extracted using 4% SDS, were partitioned into phenol and then treated with proteinase K to remove
Figure Legend Snippet: Identification within Echinacea purpurea root of 2,3-dihydroxypropyl cysteine, a structural component unique to prokaryotic lipoproteins. Lipoproteins were extracted using 4% SDS, were partitioned into phenol and then treated with proteinase K to remove

Techniques Used:

3) Product Images from "Predicted poxvirus FEN1-like nuclease required for homologous recombination, double-strand break repair and full-size genome formation"

Article Title: Predicted poxvirus FEN1-like nuclease required for homologous recombination, double-strand break repair and full-size genome formation

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.0909529106

Electrophoretic analysis of DNA. ( A ) Pulsed-field gel electrophoresis. BS-C-1 cells were infected with wt VACV or vΔG5 for the indicated times and the cell pellets were embedded in agarose plugs and lysed with SDS, EDTA and proteinase K. DNA was
Figure Legend Snippet: Electrophoretic analysis of DNA. ( A ) Pulsed-field gel electrophoresis. BS-C-1 cells were infected with wt VACV or vΔG5 for the indicated times and the cell pellets were embedded in agarose plugs and lysed with SDS, EDTA and proteinase K. DNA was

Techniques Used: Pulsed-Field Gel, Electrophoresis, Infection

Related Articles

High Performance Liquid Chromatography:

Article Title: Carbohydrate complexity structures stable diversity in gut-derived microbial consortia under high dilution pressure
Article Snippet: SCFA (acetate, propionate and butyrate) concentrations were quantified using an Agilent 7890A gas chromatograph (GC-FID 7890A, Santa Clara, CA) with a fused silica capillary column (Nukon SUPELCO No:40369-03A, Bellefonte, PA). .. SAX consortia culture at day 7 were analyzed for formate, lactate, succinate and ethanol concentrations using a HPLC system (Waters Corporation, Milford, MA) that was equipped with an organic acid column (BioRad Aminex HPX-87) and a refractive detector (Model 2414, Waters Corporation, Milford, MA). ..

Article Title: Interspecies metabolite transfer in a co-culture of Dehalococcoides and Sulfurospirillum leads to rapid and complete tetrachloroethene dechlorination
Article Snippet: Hydrogen was measured using a thermal conductivity detector (AutoSystem, Perkin Elmer, Rodgau, Germany). .. Organic acids (e.g. lactate and acetate) were analyzed by HPLC and separated on an AMINEX HPX-87H column (7.8 × 300 mm; BioRad, Munich, Germany) using 5 mM as mobile phase and at a flow rate of 0.7 mL min−1. .. Compound-specific stable isotope analysis Determination of the carbon isotope composition of the chlorinated ethenes in pure culture of S. multivorans and in co-culture of S. multivorans and D. mccartyi strain BTF08 was done using gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS; Thermo GC Trace 1320 combined with Thermo-Finnigan MAT 253 IRMS, Bremen, Germany) ( ).

Article Title: Gentiobiose and cellobiose content in fresh and fermenting cucumbers and utilization of such disaccharides by lactic acid bacteria in fermented cucumber juice medium. Gentiobiose and cellobiose content in fresh and fermenting cucumbers and utilization of such disaccharides by lactic acid bacteria in fermented cucumber juice medium
Article Snippet: A minimum of 500 µl of the supernatants were transferred into glass HPLC vials. .. Organic acids and carbohydrate concentrations were measured using a 30‐cm HPX‐87H column (Bio‐Rad Laboratories) and the HPLC method described by McFeeters ( ) with some modifications. .. The operating conditions of the UFLC Shimadzu HPLC (Shimadzu Corporation) system were a column temperature of 65°C and a 0.01 N H2 SO4 eluent at 0.9 ml/min.

Article Title: Alterations of Cellular Physiology in Escherichia coli in Response to Oxidative Phosphorylation Impaired by Defective F1-ATPase
Article Snippet: The concentration of glucose remaining in the culture broth was determined by the glucose oxidase method, using Glucose C2 (Wako Pure Chemical Industries, Ltd., Osaka, Japan). .. Organic acids in the culture broth were determined by high-pressure liquid chromatography (column, AMINEX HPX-87H; mobile phase, 0.01 N H2 SO4 ; flow rate, 0.6 ml/min; detection, absorbance at 210 nm; Bio-Rad Laboratories, Hercules, CA). ..

Spectroscopy:

Article Title: Modification of Ammonia Decomposition Activity of Ruthenium Nanoparticles by N-Doping of CNT Supports
Article Snippet: Thermogravimetric analyses of the CNT supports were carried out using a Perkin Elmer Pyris 1 TGA using approximately 2 mg of sample which is heated to 100 °C to remove any adsorbed gases and subsequently heated to 600 °C at 10 °C min−1 under a constant nitrogen flow at 20 mL min−1 . .. The organic groups in the samples were characterized by Fourier transform infrared spectroscopy (FTIR) using a Bio-Rad instrument in the range of 400–4000 cm−1 with attenuated total reflection (ATR) mode. .. Testing of Catalytic Activity Ammonia decomposition activity was tested in a catalytic bespoke flow rig using 25 mg of catalyst diluted in 450 mg of silicon carbide in a packed bed inside a quartz U-shaped reactor.

Flow Cytometry:

Article Title: Alterations of Cellular Physiology in Escherichia coli in Response to Oxidative Phosphorylation Impaired by Defective F1-ATPase
Article Snippet: The concentration of glucose remaining in the culture broth was determined by the glucose oxidase method, using Glucose C2 (Wako Pure Chemical Industries, Ltd., Osaka, Japan). .. Organic acids in the culture broth were determined by high-pressure liquid chromatography (column, AMINEX HPX-87H; mobile phase, 0.01 N H2 SO4 ; flow rate, 0.6 ml/min; detection, absorbance at 210 nm; Bio-Rad Laboratories, Hercules, CA). ..

Isolation:

Article Title: Extraterrestrial hexamethylenetetramine in meteorites—a precursor of prebiotic chemistry in the inner solar system
Article Snippet: The fraction was then frozen and dried up by a vacuum freeze dryer (EYELA Co., Ltd) under ambient temperature. .. To remove inorganic salts and interfering organic matrix from the extracts, we isolated the HMT fraction using the cation exchange chromatography (AG50W-X8 resin, Bio-Rad Laboratories) . .. The final elution containing HMTs was dried by a vacuum freeze dryer (EYELA Co., Ltd) under ambient temperature.

HMT Assay:

Article Title: Extraterrestrial hexamethylenetetramine in meteorites—a precursor of prebiotic chemistry in the inner solar system
Article Snippet: The fraction was then frozen and dried up by a vacuum freeze dryer (EYELA Co., Ltd) under ambient temperature. .. To remove inorganic salts and interfering organic matrix from the extracts, we isolated the HMT fraction using the cation exchange chromatography (AG50W-X8 resin, Bio-Rad Laboratories) . .. The final elution containing HMTs was dried by a vacuum freeze dryer (EYELA Co., Ltd) under ambient temperature.

Chromatography:

Article Title: Extraterrestrial hexamethylenetetramine in meteorites—a precursor of prebiotic chemistry in the inner solar system
Article Snippet: The fraction was then frozen and dried up by a vacuum freeze dryer (EYELA Co., Ltd) under ambient temperature. .. To remove inorganic salts and interfering organic matrix from the extracts, we isolated the HMT fraction using the cation exchange chromatography (AG50W-X8 resin, Bio-Rad Laboratories) . .. The final elution containing HMTs was dried by a vacuum freeze dryer (EYELA Co., Ltd) under ambient temperature.

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    Bio-Rad proteinase k
    Detection of glucose oxidase (GOX) and defensin-1 (Def-1) in honeydew honey samples (n = 23) and medical-grade manuka and kanuka honey following <t>proteinase</t> K treatment by immunoblotting. Aliquots (15 μl) of 50% (w/v) honey solution with or without proteinase K were resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 16.5% Tricine-SDS-PAGE. After semi-dry blotting procedure, the blocked membrane was incubated overnight with a rabbit polyclonal antibody against honeybee GOX or Def-1 (1:2000). Shown are cropped blots. (The blots with indicated cropping lines are shown in Supplementary Fig. S2 ). Immunoreactive bands were detected in solution containing dissolved SigmaFast 3,3-diaminobenzidine tablets (GOX) or detected using an enhanced chemiluminescence Immobilon Western kit (Def-1).
    Proteinase K, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Detection of glucose oxidase (GOX) and defensin-1 (Def-1) in honeydew honey samples (n = 23) and medical-grade manuka and kanuka honey following proteinase K treatment by immunoblotting. Aliquots (15 μl) of 50% (w/v) honey solution with or without proteinase K were resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 16.5% Tricine-SDS-PAGE. After semi-dry blotting procedure, the blocked membrane was incubated overnight with a rabbit polyclonal antibody against honeybee GOX or Def-1 (1:2000). Shown are cropped blots. (The blots with indicated cropping lines are shown in Supplementary Fig. S2 ). Immunoreactive bands were detected in solution containing dissolved SigmaFast 3,3-diaminobenzidine tablets (GOX) or detected using an enhanced chemiluminescence Immobilon Western kit (Def-1).

    Journal: Scientific Reports

    Article Title: Phytochemicals-mediated production of hydrogen peroxide is crucial for high antibacterial activity of honeydew honey

    doi: 10.1038/s41598-018-27449-3

    Figure Lengend Snippet: Detection of glucose oxidase (GOX) and defensin-1 (Def-1) in honeydew honey samples (n = 23) and medical-grade manuka and kanuka honey following proteinase K treatment by immunoblotting. Aliquots (15 μl) of 50% (w/v) honey solution with or without proteinase K were resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 16.5% Tricine-SDS-PAGE. After semi-dry blotting procedure, the blocked membrane was incubated overnight with a rabbit polyclonal antibody against honeybee GOX or Def-1 (1:2000). Shown are cropped blots. (The blots with indicated cropping lines are shown in Supplementary Fig. S2 ). Immunoreactive bands were detected in solution containing dissolved SigmaFast 3,3-diaminobenzidine tablets (GOX) or detected using an enhanced chemiluminescence Immobilon Western kit (Def-1).

    Article Snippet: Detection of GOX and Def-1 by immunoblotting Aliquots (15 μl) of 50% (w/v) honey solution with or without proteinase K were resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 16.5% Tricine-SDS-PAGE using a Mini-Protean II electrophoresis cell (Bio-Rad).

    Techniques: Polyacrylamide Gel Electrophoresis, SDS Page, Semi Dry Blot, Incubation, Western Blot

    Antibacterial activity of honeydew honey samples (n = 23) and medical-grade manuka and kanuka honey following catalase and proteinase K treatment against ( A ) Staphylococcus aureus and ( B ) Pseudomonas aeruginosa isolates. The 50% (w/v) honey solutions were treated with catalase (2000–5000 U/mg protein) at a final concentration ranging from 1000 to 2500 U/ml at room temperature for 2 h or proteinase K (30 U/mg) at a final concentration of 50 μg/ml at 37 °C for 30 min. The antibacterial activity was determined with a minimum inhibitory concentration (MIC) assay. The MIC was defined as the lowest concentration of honey solution (%) inhibiting bacterial growth. K, kanuka honey; M, manuka honey.

    Journal: Scientific Reports

    Article Title: Phytochemicals-mediated production of hydrogen peroxide is crucial for high antibacterial activity of honeydew honey

    doi: 10.1038/s41598-018-27449-3

    Figure Lengend Snippet: Antibacterial activity of honeydew honey samples (n = 23) and medical-grade manuka and kanuka honey following catalase and proteinase K treatment against ( A ) Staphylococcus aureus and ( B ) Pseudomonas aeruginosa isolates. The 50% (w/v) honey solutions were treated with catalase (2000–5000 U/mg protein) at a final concentration ranging from 1000 to 2500 U/ml at room temperature for 2 h or proteinase K (30 U/mg) at a final concentration of 50 μg/ml at 37 °C for 30 min. The antibacterial activity was determined with a minimum inhibitory concentration (MIC) assay. The MIC was defined as the lowest concentration of honey solution (%) inhibiting bacterial growth. K, kanuka honey; M, manuka honey.

    Article Snippet: Detection of GOX and Def-1 by immunoblotting Aliquots (15 μl) of 50% (w/v) honey solution with or without proteinase K were resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 16.5% Tricine-SDS-PAGE using a Mini-Protean II electrophoresis cell (Bio-Rad).

    Techniques: Activity Assay, Concentration Assay

    MMS produces heat-labile DNA damage in S.cerevisiae . PFGE of yeast chromosomes after a 0.5 h MMS treatment (0.05%) and treatment with proteinase K at 50°C ( A ) or at 30°C ( B ) for 24 h. Chromosomes were visualized by ethidium bromide or Southern hybridization to highlight chromosome VIII directly after MMS treatments or following repair as indicated.

    Journal: Nucleic Acids Research

    Article Title: Methyl methanesulfonate (MMS) produces heat-labile DNA damage but no detectable in vivo DNA double-strand breaks

    doi: 10.1093/nar/gki681

    Figure Lengend Snippet: MMS produces heat-labile DNA damage in S.cerevisiae . PFGE of yeast chromosomes after a 0.5 h MMS treatment (0.05%) and treatment with proteinase K at 50°C ( A ) or at 30°C ( B ) for 24 h. Chromosomes were visualized by ethidium bromide or Southern hybridization to highlight chromosome VIII directly after MMS treatments or following repair as indicated.

    Article Snippet: Inserts were incubated in 0.5 M EDTA, 1% N -laurylsarcosyl and proteinase K (1 mg/ml) at 50 or 20°C for 48 h, and thereafter washed four times in TE-buffer prior to loading onto an agarose separation gel (1% Chromosomal grade agarose, Bio-Rad).

    Techniques: Hybridization

    The pro-inflammatory response of HEp-2 cells to A. baumannii OMVs treated with proteinase K and EDTA. (A) Protein profiles of A. baumannii OMVs. Lane M, size marker; 1, purified intact OMVs; 2, OMVs treated with EDTA; 3, OMVs treated with proteinase K; 4, proteinase K. (B) Expression of pro-inflammatory cytokine genes to proteinase K- and EDTA-treated A. baumannii OMVs was assessed by quantitative real-time PCR. HEp-2 cells were treated with the same concentration (15 µg/ml) of A. baumannii OMVs for 24 h as a positive control. Data are presented as mean ± SD of duplicate determinations.

    Journal: PLoS ONE

    Article Title: Acinetobacter baumannii Outer Membrane Vesicles Elicit a Potent Innate Immune Response via Membrane Proteins

    doi: 10.1371/journal.pone.0071751

    Figure Lengend Snippet: The pro-inflammatory response of HEp-2 cells to A. baumannii OMVs treated with proteinase K and EDTA. (A) Protein profiles of A. baumannii OMVs. Lane M, size marker; 1, purified intact OMVs; 2, OMVs treated with EDTA; 3, OMVs treated with proteinase K; 4, proteinase K. (B) Expression of pro-inflammatory cytokine genes to proteinase K- and EDTA-treated A. baumannii OMVs was assessed by quantitative real-time PCR. HEp-2 cells were treated with the same concentration (15 µg/ml) of A. baumannii OMVs for 24 h as a positive control. Data are presented as mean ± SD of duplicate determinations.

    Article Snippet: OMV samples, including intact OMVs and proteinase K- and EDTA-treated OMVs, were separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with Coomassie brilliant blue R-250 (Bio-Rad).

    Techniques: Marker, Purification, Expressing, Real-time Polymerase Chain Reaction, Concentration Assay, Positive Control

    PrP Sc typing of sheep scrapie and human sporadic CJD. After treatment with proteinase K alone or combined with PNGase F, Western blot analysis of proteinase K digested classical scrapie cases ( A ) showed the typical triplet pattern of PrP Sc , whereas atypical/Nor98

    Journal: The American Journal of Pathology

    Article Title: Similarities between Forms of Sheep Scrapie and Creutzfeldt-Jakob Disease Are Encoded by Distinct Prion Types

    doi: 10.2353/ajpath.2009.090623

    Figure Lengend Snippet: PrP Sc typing of sheep scrapie and human sporadic CJD. After treatment with proteinase K alone or combined with PNGase F, Western blot analysis of proteinase K digested classical scrapie cases ( A ) showed the typical triplet pattern of PrP Sc , whereas atypical/Nor98

    Article Snippet: On treatment with proteinase K (see above) samples were diluted in Tris-buffered saline and subjected to a commercially available slot blot/dot blot device (Bio-Rad), run by a diaphragm pump.

    Techniques: Western Blot