thermolabile proteinase k  (New England Biolabs)


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    Name:
    Thermolabile Proteinase K
    Description:
    Thermolabile Proteinase K is an engineered subtilisin related serine protease that cleaves the peptide bond at the carboxyl side of aliphatic or aromatic amino acid residues and will hydrolyze a variety of peptide bonds thus degrading unwanted proteins Unlike native Proteinase K this variation of the enzyme is inactivated when heated for 10 minutes or greater at 55ºC
    Catalog Number:
    P8111S
    Price:
    153
    Category:
    Proteases
    Size:
    30 units
    Buy from Supplier


    Structured Review

    New England Biolabs thermolabile proteinase k
    Thermolabile Proteinase K
    Thermolabile Proteinase K is an engineered subtilisin related serine protease that cleaves the peptide bond at the carboxyl side of aliphatic or aromatic amino acid residues and will hydrolyze a variety of peptide bonds thus degrading unwanted proteins Unlike native Proteinase K this variation of the enzyme is inactivated when heated for 10 minutes or greater at 55ºC
    https://www.bioz.com/result/thermolabile proteinase k/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    thermolabile proteinase k - by Bioz Stars, 2021-06
    96/100 stars

    Images

    1) Product Images from "Preparation of E. coli RNA polymerase transcription elongation complexes for systematic RNA assays"

    Article Title: Preparation of E. coli RNA polymerase transcription elongation complexes for systematic RNA assays

    Journal: bioRxiv

    doi: 10.1101/2021.03.15.435517

    Integration of TEC purification with USER barcoding. (A) Degradation of purified TECs using Thermolabile Proteinase K (TL Prot. K) followed by heat treatment at 65 °C. (B) Stepwise visualization of the USER barcoding procedure beginning from a single pool of purified TECs.
    Figure Legend Snippet: Integration of TEC purification with USER barcoding. (A) Degradation of purified TECs using Thermolabile Proteinase K (TL Prot. K) followed by heat treatment at 65 °C. (B) Stepwise visualization of the USER barcoding procedure beginning from a single pool of purified TECs.

    Techniques Used: Purification

    2) Product Images from "Preparation of E. coli RNA polymerase transcription elongation complexes for systematic RNA assays"

    Article Title: Preparation of E. coli RNA polymerase transcription elongation complexes for systematic RNA assays

    Journal: bioRxiv

    doi: 10.1101/2021.03.15.435517

    Integration of TEC purification with USER barcoding. (A) Degradation of purified TECs using Thermolabile Proteinase K (TL Prot. K) followed by heat treatment at 65 °C. (B) Stepwise visualization of the USER barcoding procedure beginning from a single pool of purified TECs.
    Figure Legend Snippet: Integration of TEC purification with USER barcoding. (A) Degradation of purified TECs using Thermolabile Proteinase K (TL Prot. K) followed by heat treatment at 65 °C. (B) Stepwise visualization of the USER barcoding procedure beginning from a single pool of purified TECs.

    Techniques Used: Purification

    3) Product Images from "Direct on-the-spot detection of SARS-CoV-2 in patients"

    Article Title: Direct on-the-spot detection of SARS-CoV-2 in patients

    Journal: Experimental Biology and Medicine

    doi: 10.1177/1535370220941819

    Protocol adjustment and optimal conditions. (a) Schematic representation of the isothermal colorimetric RT-LAMP reaction. (b) RT-LAMP reaction performed on purified RNA from nasal and throat swabs submerged in UTM buffer. Results from a no-template control (NTC; left), a negative subject (Neg S.; middle), and a positive subject (Pos S.; right) at t = 0 (upper panels) and after a 30-min incubation at 65°C (lower panels) are shown. Three technical replicates of each sample are shown. (c) Representative RT-LAMP test results of clinical diagnostic nasal and throat swabs. Three different negative samples (Neg S.; left), and three different positive samples (Pos S.; right) obtained at t = 0 and t = 30 min were directly tested with no RNA purification step. (d) Comparison of the RT-LAMP method to Ct values obtained by standard RT-qPCR (3 true positive and 2 false negative samples of the 99 samples analyzed are not shown due to inaccessibility to their RT-qPCR Ct values). RT-qPCR negative samples were assigned arbitrary Ct values, for visualization. (e) Classification of true positive (TP), true negative (TN), false positive (FP) and false negative (FN) numbers and rate of RT-LAMP test results, as compared to the standard RT-qPCR test results. (f) Clinical diagnostic nasal and throat swabs tested by two different RT-LAMP protocols. Upper panel, without proteinase K and guanidine hydrochloride. Lower panel, with proteinase K and guanidine hydrochloride. For RT-qPCR positive samples, the Ct value is presented under each sample. The sample to the right is negative. (g) RT-LAMP results for samples from patients confirmed as positive for the following viruses, with the fraction tested indicated: 1–2, HSV; swabs (lysed and inactivated as described in the currently developed protocol). 3, HSV; purified DNA. 4, RSV; purified RNA. 5, Influenza B; RNA. 6, Enterovirus; RNA. 7, RNA extraction from a SARS-CoV-2 positive patient. 8, no template control. Results are shown at t = 0 and 30 min after incubation at 65 ͦC.
    Figure Legend Snippet: Protocol adjustment and optimal conditions. (a) Schematic representation of the isothermal colorimetric RT-LAMP reaction. (b) RT-LAMP reaction performed on purified RNA from nasal and throat swabs submerged in UTM buffer. Results from a no-template control (NTC; left), a negative subject (Neg S.; middle), and a positive subject (Pos S.; right) at t = 0 (upper panels) and after a 30-min incubation at 65°C (lower panels) are shown. Three technical replicates of each sample are shown. (c) Representative RT-LAMP test results of clinical diagnostic nasal and throat swabs. Three different negative samples (Neg S.; left), and three different positive samples (Pos S.; right) obtained at t = 0 and t = 30 min were directly tested with no RNA purification step. (d) Comparison of the RT-LAMP method to Ct values obtained by standard RT-qPCR (3 true positive and 2 false negative samples of the 99 samples analyzed are not shown due to inaccessibility to their RT-qPCR Ct values). RT-qPCR negative samples were assigned arbitrary Ct values, for visualization. (e) Classification of true positive (TP), true negative (TN), false positive (FP) and false negative (FN) numbers and rate of RT-LAMP test results, as compared to the standard RT-qPCR test results. (f) Clinical diagnostic nasal and throat swabs tested by two different RT-LAMP protocols. Upper panel, without proteinase K and guanidine hydrochloride. Lower panel, with proteinase K and guanidine hydrochloride. For RT-qPCR positive samples, the Ct value is presented under each sample. The sample to the right is negative. (g) RT-LAMP results for samples from patients confirmed as positive for the following viruses, with the fraction tested indicated: 1–2, HSV; swabs (lysed and inactivated as described in the currently developed protocol). 3, HSV; purified DNA. 4, RSV; purified RNA. 5, Influenza B; RNA. 6, Enterovirus; RNA. 7, RNA extraction from a SARS-CoV-2 positive patient. 8, no template control. Results are shown at t = 0 and 30 min after incubation at 65 ͦC.

    Techniques Used: Purification, Incubation, Diagnostic Assay, Quantitative RT-PCR, RNA Extraction

    Related Articles

    other:

    Article Title: TCF7L2 regulates postmitotic differentiation programs and excitability patterns in the thalamus
    Article Snippet: Cross-links were reversed and both eluates and input controls were treated with RNAse and Proteinase K, according to ( ).

    Article Title: Preparation of E. coli RNA polymerase transcription elongation complexes for systematic RNA assays
    Article Snippet: Proteins All enzymes, including Q5® High-Fidelity DNA Polymerase, Q5U® High-Fidelity DNA Polymerase, Vent® (exo-) DNA polymerase, Sulfolobus DNA Polymerase IV, Thermolabile Exonuclease I, Klenow Fragment (3’→5’ exo-), Klenow Fragment, T4 DNA Polymerase, Thermolabile USER® II Enzyme, T4 DNA Ligase, E. coli RNA Polymerase holoenzyme, Thermolabile Proteinase K, and RNase If were purchased from New England Biolabs (NEB).

    Blocking Assay:

    Article Title: Direct on-the-spot detection of SARS-CoV-2 in patients
    Article Snippet: For calibration, reactions were performed with or without proteinase K and/or guanidine hydrochloride. .. The proteinase K inactivation and reaction steps were performed in a closed-lid heating block or, for proof of concept, in a Thermos cup, using warm water. ..

    Lysis:

    Article Title: Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction
    Article Snippet: .. Lysis buffer consisted of 0.1-fold buffer TE pH 8.0 (Ambion, AM9848) with 0.1% TWEEN-20, 1% volume (e.g., 1 µL enzyme added to 100µL buffer) Thermolabile Proteinase K (NEB, P8111S), 2% volume ezDNase (Invitrogen, 11766051), and 0.3 ng/µL human genomic DNA from a normal male. .. For one reaction, 460 µL of reaction mix and 20 µL of lysis buffer were preloaded in a clean 1.5 mL DNA LoBind microcentrifuge tube (Eppendorf, 022431021) and kept on ice until use.

    Purification:

    Article Title: STAG2 loss-of-function affects short-range genomic contacts and modulates urothelial differentiation in bladder cancer cells
    Article Snippet: DNA was recovered in elution buffer (1% SDS, 0.1M NaHCO3 ) and cross-linking was reversed by overnight incubation at 65 °C. .. RNA and proteins were sequentially digested with 20 μ g of RNAse and 40 μg of proteinase K. DNA was purified by phenol-chloroform extraction and resuspended in TE 0.5X. ..

    Incubation:

    Article Title: Preparation of E. coli RNA polymerase transcription elongation complexes for systematic RNA assays
    Article Snippet: A 15 μl aliquot was removed and assessed by EMSA as described above in the section Analysis of purified TECs by EMSA , and a 25 μl ‘Input’ fraction was mixed with 125 μl of Stop Solution, phenol:chloform extracted and set up for ethanol precipitation as described above in the section P RA1 barcoding validation assays . .. 150 μl of the remaining sample was mixed with 6 μl (1 μl per sample volume) of Thermolabile Proteinase K (NEB), incubated at 37 °C for 30 minutes, and then at 65 °C for 20 minutes. .. 3 μl (0.5 μl per sample volume) of RNase If (NEB) was added and the sample was incubated at 37 °C for 15 minutes, and then at 70 °C for 20 minutes.

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  • 96
    New England Biolabs thermolabile proteinase k
    Integration of TEC purification with USER barcoding. (A) Degradation of purified TECs using Thermolabile <t>Proteinase</t> K (TL Prot. K) followed by heat treatment at 65 °C. (B) Stepwise visualization of the USER barcoding procedure beginning from a single pool of purified TECs.
    Thermolabile Proteinase K, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/thermolabile proteinase k/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    thermolabile proteinase k - by Bioz Stars, 2021-06
    96/100 stars
      Buy from Supplier

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    Integration of TEC purification with USER barcoding. (A) Degradation of purified TECs using Thermolabile Proteinase K (TL Prot. K) followed by heat treatment at 65 °C. (B) Stepwise visualization of the USER barcoding procedure beginning from a single pool of purified TECs.

    Journal: bioRxiv

    Article Title: Preparation of E. coli RNA polymerase transcription elongation complexes for systematic RNA assays

    doi: 10.1101/2021.03.15.435517

    Figure Lengend Snippet: Integration of TEC purification with USER barcoding. (A) Degradation of purified TECs using Thermolabile Proteinase K (TL Prot. K) followed by heat treatment at 65 °C. (B) Stepwise visualization of the USER barcoding procedure beginning from a single pool of purified TECs.

    Article Snippet: 150 μl of the remaining sample was mixed with 6 μl (1 μl per sample volume) of Thermolabile Proteinase K (NEB), incubated at 37 °C for 30 minutes, and then at 65 °C for 20 minutes.

    Techniques: Purification

    Protocol adjustment and optimal conditions. (a) Schematic representation of the isothermal colorimetric RT-LAMP reaction. (b) RT-LAMP reaction performed on purified RNA from nasal and throat swabs submerged in UTM buffer. Results from a no-template control (NTC; left), a negative subject (Neg S.; middle), and a positive subject (Pos S.; right) at t = 0 (upper panels) and after a 30-min incubation at 65°C (lower panels) are shown. Three technical replicates of each sample are shown. (c) Representative RT-LAMP test results of clinical diagnostic nasal and throat swabs. Three different negative samples (Neg S.; left), and three different positive samples (Pos S.; right) obtained at t = 0 and t = 30 min were directly tested with no RNA purification step. (d) Comparison of the RT-LAMP method to Ct values obtained by standard RT-qPCR (3 true positive and 2 false negative samples of the 99 samples analyzed are not shown due to inaccessibility to their RT-qPCR Ct values). RT-qPCR negative samples were assigned arbitrary Ct values, for visualization. (e) Classification of true positive (TP), true negative (TN), false positive (FP) and false negative (FN) numbers and rate of RT-LAMP test results, as compared to the standard RT-qPCR test results. (f) Clinical diagnostic nasal and throat swabs tested by two different RT-LAMP protocols. Upper panel, without proteinase K and guanidine hydrochloride. Lower panel, with proteinase K and guanidine hydrochloride. For RT-qPCR positive samples, the Ct value is presented under each sample. The sample to the right is negative. (g) RT-LAMP results for samples from patients confirmed as positive for the following viruses, with the fraction tested indicated: 1–2, HSV; swabs (lysed and inactivated as described in the currently developed protocol). 3, HSV; purified DNA. 4, RSV; purified RNA. 5, Influenza B; RNA. 6, Enterovirus; RNA. 7, RNA extraction from a SARS-CoV-2 positive patient. 8, no template control. Results are shown at t = 0 and 30 min after incubation at 65 ͦC.

    Journal: Experimental Biology and Medicine

    Article Title: Direct on-the-spot detection of SARS-CoV-2 in patients

    doi: 10.1177/1535370220941819

    Figure Lengend Snippet: Protocol adjustment and optimal conditions. (a) Schematic representation of the isothermal colorimetric RT-LAMP reaction. (b) RT-LAMP reaction performed on purified RNA from nasal and throat swabs submerged in UTM buffer. Results from a no-template control (NTC; left), a negative subject (Neg S.; middle), and a positive subject (Pos S.; right) at t = 0 (upper panels) and after a 30-min incubation at 65°C (lower panels) are shown. Three technical replicates of each sample are shown. (c) Representative RT-LAMP test results of clinical diagnostic nasal and throat swabs. Three different negative samples (Neg S.; left), and three different positive samples (Pos S.; right) obtained at t = 0 and t = 30 min were directly tested with no RNA purification step. (d) Comparison of the RT-LAMP method to Ct values obtained by standard RT-qPCR (3 true positive and 2 false negative samples of the 99 samples analyzed are not shown due to inaccessibility to their RT-qPCR Ct values). RT-qPCR negative samples were assigned arbitrary Ct values, for visualization. (e) Classification of true positive (TP), true negative (TN), false positive (FP) and false negative (FN) numbers and rate of RT-LAMP test results, as compared to the standard RT-qPCR test results. (f) Clinical diagnostic nasal and throat swabs tested by two different RT-LAMP protocols. Upper panel, without proteinase K and guanidine hydrochloride. Lower panel, with proteinase K and guanidine hydrochloride. For RT-qPCR positive samples, the Ct value is presented under each sample. The sample to the right is negative. (g) RT-LAMP results for samples from patients confirmed as positive for the following viruses, with the fraction tested indicated: 1–2, HSV; swabs (lysed and inactivated as described in the currently developed protocol). 3, HSV; purified DNA. 4, RSV; purified RNA. 5, Influenza B; RNA. 6, Enterovirus; RNA. 7, RNA extraction from a SARS-CoV-2 positive patient. 8, no template control. Results are shown at t = 0 and 30 min after incubation at 65 ͦC.

    Article Snippet: The proteinase K inactivation and reaction steps were performed in a closed-lid heating block or, for proof of concept, in a Thermos cup, using warm water.

    Techniques: Purification, Incubation, Diagnostic Assay, Quantitative RT-PCR, RNA Extraction