Proteinase k neb. New England Biolabs GmbH Brüningstr.
Proteinase k neb FAQ: How do I heat inactivate Thermolabile Proteinase K? For the majority of workflows, Thermolabile Proteinase K can be heat inactivated by incubating at 55°C for 10 FAQ: What is the Proteinase K activity in commonly used buffers? Buffer: 30 mM Tris HCl, pH 8. Links to this resource Related Products: Thermolabile P8107 - Proteinase K Revision date 29-Aug-2023 Methods and material for containment and cleaning up Methods for containment Prevent further leakage or spillage if safe to do so. It preferentially cleaves the peptide bond at the carboxyl side of FAQ: What is the activity if Proteinase K, Molecular Biology Grade if subjected to the unit definition assay used for Thermolabile Proteinase K? Proteinase K, Molecular Biology Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. Furin (NEB #P8077) is a ubiquitous subtilisin-like proprotein convertase with a What buffer should I use for Proteinase K? If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this? Why is the unit definition assay for Thermolabile Proteinase K NEB offers several proteases with different recognition sites. Ph. 05 to 1 mg/mL. Effective March 14, 2024: Proteinase K, Molecular Biology Grade component number has changed with the concentration changing from 800U/ml to 20 mg/ml. Engineered for Proteinase K from the fungus Engyodontium album is a nonspecific serine protease that is useful for general digestion of proteins. Thermostable RNase H can be inactivated by treatment with proteinase K or addition of excess EDTA. Home Reagents and Proteinase K, Molecular Biology Grade TEV Protease Thermolabile Proteinase K Trypsin-ultra™, Mass Proteinase K NEB is a true enzymatic masterpiece, its proteolytic prowess enabling researchers to explore the intricate world of DNA, proteins, and cellular processes. Proteinase K is a proteolytic enzyme (a serine protease) that is purified from the Using Thermolabile Proteinase K allows one to avoid performing phenol/chloroform extraction or gel/column purification. 2. This is a good starting point; however, the Proteinase K is a broad-spectrum serine protease and our product is extracted from Pichia pastoris cells with cloned gene encoding Engyodontium album (Tritirachium album) endolytic Effective March 14, 2024: Proteinase K, Molecular Biology Grade component number has changed with the concentration changing from 800U/ml to 20 mg/ml. Contact your local UK NEBExpress Salt Active Nuclease can be degraded with Proteinase K , or Thermolabile Proteinase K . Proteinase K は、全てのNEB制限酵素バッファーを含む幅広いバッファー中で活性を示す。pH 7. com Emergency telephone number Emergency telephone 978-927-5054 800-632-5227 (toll free) Product name Monarch Proteinase K Resuspension Buffer FAQ: What buffer should I use for Proteinase K? Proteinase K is active in a wide range of buffers including all Restriction Enzyme NEBuffers, Q5 Reaction Buffer, OneTaq Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. Incubate for 60 Product name Proteinase K Product No P8107 Other means of identification Synonyms None E-mail address info. Links to this resource Related For Questions Related E-mail address info@neb. What is the protein concentration of Proteinase K, Molecular Biology Grade? What buffer should I use for Recombinant Proteinase K, heat inactivated completely at 55°C in 10 min, with broad protease activity. HAZARDS Proteinase K, Molecular Biology Grade Product information. Contact your local US Buffer and 20 µl of Proteinase K for 45 minutes at 56°C in a thermal mixer with agitation set at 300 rpm. It preferentially cleaves the peptide bond at the carboxyl side of FAQ: How do I heat inactivate Thermolabile Proteinase K? For the majority of workflows, Thermolabile Proteinase K can be heat inactivated by incubating at 55°C for 10 FAQ: What is the optimal reaction buffer for Thermolabile Proteinase K? Thermolabile Proteinase K is compatible with a wide range of buffers: Optimal pH: 7–10 Salt: However, many Proteinase K products do not have the same specifications as NEB’s Proteinase K , which has been highly characterized for more consistent performance. This enzyme is widely used to remove proteins/enzymes in nucleic After the RNA from the cytoplasm is digested by RNase A, the Nuclei Lysis Buffer, pre-mixed with the included Proteinase K, is used to lyse the nuclei and release the HMW genomic DNA. com 24 Hour Emergency Phone Number Chemtrec +1 703-741-5970 Product name Proteinase K Page 1 / 9 Product No P8107 Supplier Address New Product For most applications, the same volume of Thermolabile Proteinase K can be used in place of Proteinase K, Molecular Biology Grade. au@neb. FAQ: What is the Proteinase K activity in commonly used buffers? Buffer: 30 mM Tris HCl, pH 8. New England Biolabs GmbH Brüningstr. 0 µmol of 4 For the group with Proteinase K digestion, virions were incubated with 0. com 24 Hour Emergency Phone Number Chemtrec +1 703-741-5970 Page 1 / 10 Supplier Address New England BioLabs 240 County Road Ipswich, MA 01938 Thermolabile Proteinase K Close Trypsin-ultra™, Mass Spectrometry Grade Close α2-3 Neuraminidase S Close α-Lytic Protease If you have any questions, please contact Proteinase K from the fungus Engyodontium album is a nonspecific serine protease that is useful for general digestion of proteins. 0–12. Proteinase K is active in a wide range Proteinase K, RNase A and Blood Lysis Buffer to the frozen sample in step 2. Rinse mouth. Its For immediate use, Proteinase K can be diluted into 1X reaction buffer. support phone: +1(800)632-7799 Monarch Proteinase K (20. 1 New England Biolabs New England Biolabs New England Biolabs (NEB) Tech. • The recommended working concentration of Proteinase K is 0. After lysis, 300 µl Protein Separation Solution was added, gently mixed, and centri-fuged Thermolabile Proteinase K NEB P8111 New England Biolabs. Just updated. Never give anything Proteinase K is a proteolytic enzyme (a serine protease) that is purified from the mold Tritirachium album. 5 ml) Monarch Proteinase K Reaction Buffer (4 ml) Monarch DNase I (275 units) Monarch DNase I Reaction For Proteinase K treated samples, doubling Proteinase K (from 5% to 10%) may lead to an increase in RNA yield Too much sample: Reduce amount of starting material to match kit FAQ: Can I use leftover Proteinase K and DNase I from the Monarch ® Total RNA Miniprep Kit for other applications? Yes, but the formulations have been optimized for use with Product name Proteinase K Product No P8107 Other means of identification Synonyms None E-mail address info. Our website shows product prices Sign In or Sign UpMy NEB. com Thermostable Endonuclease Q (EndoQ) is an archaeal DNA endonuclease that cleaves the phosphate backbone of a DNA substrate 5′ to the position of the modified Using Thermolabile Proteinase K allows one to avoid performing phenol/chloroform extraction or gel/column purification. Proteinase K, Molecular Biology Grade. Links to this resource Product Categories: Proteases Products For Questions Related to NEB Products and Mechanical homogenization prior to digestion with Proteinase K often increases RNA yield. It preferentially cleaves the peptide bond at the carboxyl side of Product Name: Thermolabile Proteinase K Catalog #: P8111S Concentration: 120 units/ml Unit Definition: One unit is defined as the amount of enzyme required to release 1. Doubling the amount of Proteinase K and/or increasing incubation time may also increase A unique, recombinant Proteinase K that retains similar activity and specificity as wild type but can be completely heat inactivated at lower temperatures compared to others. After incubation for 16 hours, >90% of the substrate RNA remains intact as determined by gel P8107 - Proteinase K Revision date 29-Aug-2023 Methods and material for containment and cleaning up Methods for containment Prevent further leakage or spillage if safe to do so. Proteinase K is active in a wide range The protein concentration of Proteinase K, Molecular Biology Grade is ~20 mg/ml and its unit activity is 800 U/ml. com Emergency telephone The protein concentration of Proteinase K, Molecular Biology Grade is ~20 mg/ml and its unit activity is 800 U/ml. 0及び20～60°Cで高い活性を示す（1,2）。Proteinase K はEDTAなどのキレート剤 Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. . Did you Includes Proteinase K for processing of tissues and blood samples Can I use NEB Proteinase K Molecular Biology Grade NEB P8107 in conjunction with the Monarch Total RNA NEB offers several proteases with different recognition sites. Incubate reaction at 37˚C for 15 minutes. Proteinase K remains active: • Over a wide pH range—optimal activity between 6. Supplier: New England Biolabs, Inc. ): 100% Buffer: 50mM Tris Proteinase K, originally from the fungus Tritirachium album, is a highly active serine protease with broad cleavage specificity. First-aid measures Description of first aid measures Inhalation Remove to fresh air. Furin is a ubiquitous NEB Proteinase K is a vital tool in molecular biology, with its broad range of applications and benefits making it indispensable for researchers. Thermolabile Proteinase K is fully active in the presence of 10 mM EDTA, FAQ: Why is the unit definition assay for Thermolabile Proteinase K (NEB #P8111) different from the unit definition assay for Proteinase K, Molecular Biology Grade (NEB # To Request Technical Support. However, typical reaction FAQ: What buffer should I use for Proteinase K? Proteinase K is active in a wide range of buffers including all Restriction Enzyme NEBuffers, Q5 Reaction Buffer, OneTaq FAQ: How do I heat inactivate Thermolabile Proteinase K? For the majority of workflows, Thermolabile Proteinase K can be heat inactivated by incubating at 55°C for 10 What is the Proteinase K activity in commonly used buffers? Protocols for Protein Digestion O-Glycosidase Application Note 1 (P0733) O-Glycosidase (P0733) Endo-α-N Inc (NEB). One unit of NEBExpress Salt Active Nuclease will digest 30 µg of calf thymus Removal of reactants is also beneficial following standard protocols such as RNA labeling, capping, Proteinase K treatment, and DNase I treatment. 5 ug of a DNA/RNA modifying enzyme FAQ: What buffer should I use to dilute Thermolabile Proteinase K? The amount of Thermolabile Proteinase K required may be substrate dependent. 5 and 9. Contact your local US Sales Representative. 5 with an optimum of pH 8. de@neb. This is a good starting point; however, the E-mail address info@neb. Thermostable RNase H displays 3 FAQ: Is Thermolabile Proteinase K active in common NEB buffers? Yes, Thermolabile Proteinase K is active in NEBuffer 1. The activity of the enzyme is stimulated by 0. 15-11-2024. Expand menu. com Proteinase K (NEB #P8102) is a non-specific, subtilisin-related serine protease with a very high specific activity. Your credentials are incorrect or you are trying to login with a non-existing webshop account. 8 units of Proteinase K, Molecular Biology Grade is incubated at 37ºC. 00 What buffer should I use for Proteinase K? If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this? Why is the unit definition assay for Thermolabile Proteinase K (NEB #P8111) different from the unit definition What buffer should I use for Proteinase K? If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this? Why is the unit definition assay for Thermolabile Proteinase K (NEB #P8111) different from the unit definition FAQ: What buffer should I use for Proteinase K? Proteinase K is active in a wide range of buffers including all Restriction Enzyme NEBuffers, Q5 Reaction Buffer, OneTaq Thermolabile Proteinase K Close Trypsin-ultra™, Mass Spectrometry Grade Close α-Lytic Protease Submit If you have any questions, please contact Customer Support at FAQ: What is the Proteinase K activity in commonly used buffers? Buffer: 30 mM Tris HCl, pH 8. NEBinspired Blog . Thermolabile Proteinase K is an engineered and recombinantly expressed variant of Proteinase K, Molecular Biology Grade (wild type). It preferentially cleaves the peptide bond at the carboxyl side of For a 10-50 uL reaction, containing 40 ng (0. 00 $45. The P8107 - Proteinase K Revision date 29-Aug-2023 Methods and material for containment and cleaning up Methods for containment Prevent further leakage or spillage if safe to do so. Proteinase K is a subtilisin-related serine protease that hydrolyzes a variety of peptide bonds and is frequently used to cleanup enzymatic reactions or cell lysates. com Emergency telephone number Company Phone Number 1800 934 218 24 Hour Emergency Phone Number Chemtrec +65 3163 8374 Australian Poisons Add 1 μl of Thermolabile Proteinase K, mix gently. It preferentially cleaves the peptide bond at the carboxyl side of Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. Custom Proteinase K (20 mg/ml) properties of Thermolabile Proteinase K can be found in detail in the FAQ. Furin is a ubiquitous May 16, 2020 Thermolabile Proteinase K Typical Reaction Protocol V. Our website shows product prices Can I use leftover Proteinase K and DNase I from the Monarch® Total RNA Miniprep Kit for other applications? Can I use NEB Proteinase K, Molecular Biology Grade (NEB #P8107S) in conjunction with the Monarch® Total RNA NEB offers several proteases with different recognition sites. Its ability to work under challenging conditions and its role in preparing clean, . What is the protein concentration of Proteinase K, Molecular Biology Grade? The protein What buffer should I use for Proteinase K? If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this? Why is the unit definition assay for Thermolabile Proteinase K FAQ: What is the optimal reaction buffer for Thermolabile Proteinase K? Thermolabile Proteinase K is compatible with a wide range of buffers: Optimal pH: 7–10 Salt: FAQ: Is Thermolabile Proteinase K active in common NEB buffers? Yes, Thermolabile Proteinase K is active in NEBuffer 1. Add 10 µl Proteinase K, 3 µl RNase A and 100 µl of Blood Lysis Buffer. 1600 units ( 800 units/ml ) - (NEB E-mail address info. Genomic DNA (gDNA) is Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. P8107 - Proteinase K Revision date 29-Aug-2023 Methods and material for containment and cleaning up Methods for containment Prevent further leakage or spillage if safe to do so. D, in RNA Methodologies (Fourth Edition), 2010 Proteinase K. 2 to 1% SDS or by 1 to 4 M urea • Ca2+ protects Proteinase K against autolysis, increases the thermal stability, FAQ: Why is the unit definition assay for Thermolabile Proteinase K (NEB #P8111) different from the unit definition assay for Proteinase K, Molecular Biology Grade (NEB # Thermolabile Proteinase K is an engineered and recombinantly expressed variant of Proteinase K, Molecular Biology Grade (wild type). Tissue: Tissue pieces are too large: Cut starting material to the smallest possible pieces or Highly characterized for more consistent performance, Proteinase K is a subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. FAQ: How do I heat inactivate Thermolabile Proteinase K? For the majority of workflows, Thermolabile Proteinase K can be heat inactivated by incubating at 55°C for 10 Thermolabile Proteinase K is free of exo- and endonuclease activity, RNase activity, and contains no detectable DNA (both bacterial and eukaryotic) contamination. 5 • Under E-mail address info@neb. com Emergency telephone number Emergency telephone 978-927-5054 800-632-5227 (toll free) 9:00am - 5:00pm Monday-Friday EST 2. Do NOT induce vomiting. E-mail address info@neb. Links to this resource For Questions Related to NEB FAQ: What buffer should I use for Proteinase K? Proteinase K is active in a wide range of buffers including all Restriction Enzyme NEBuffers, Q5 Reaction Buffer, OneTaq FAQ: What buffer should I use for Proteinase K? Proteinase K is active in a wide range of buffers including all Restriction Enzyme NEBuffers, Q5 Reaction Buffer, OneTaq NEBExpress Salt Active Nuclease can be degraded with Proteinase K , or Thermolabile Proteinase K . Search. 0 Application: Reference Relative activity (approx. It preferentially cleaves the peptide bond at the carboxyl side of However, many Proteinase K products do not have the same specifications as NEB’s Proteinase K , which has been highly characterized for more consistent performance. com 24 Hour Emergency Phone Number Chemtrec +1 703-741-5970 Page 1 / 10 Supplier Address New England BioLabs 240 County Road Ipswich, MA 01938 Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. B852 D-65926 Frankfurt am Main Tel: +49-69/305-23140 E-Mail: info. ): 100% Buffer: 50mM Tris What buffer should I use for Proteinase K? If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this? Why is the unit definition assay for Thermolabile Proteinase K What buffer should I use for Proteinase K? If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this? Why is the unit definition assay for Thermolabile Proteinase K (NEB #P8111) different from the unit definition What buffer should I use for Proteinase K? If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this? Why is the unit definition assay for Thermolabile Proteinase K FAQ: The protocol I am using requires a µg/ml concentration of Proteinase K. 1, 2. However, typical reaction FAQ: There is a precipitate in the Proteinase K enzyme tube, is this normal? It is not uncommon to see a white, amorphous precipitate in the Proteinase K tube. Proteinase K is a non-specific, subtilisin-related serine protease with a very high specific activity. It preferentially cleaves the peptide bond at the carboxyl side of NEB offers several proteases with different recognition sites. 2 mg/mL Proteinase K (NEB #P8107S) in 120 μL PBS at 37°C for 30 min and washed with PBST buffer three times. Robert E. Incubate reaction at 55˚C for 10 minutes to inactivate Thermolabile Proteinase K. The cleavage patterns of both enzymes FAQ: There is a precipitate in the Proteinase K enzyme tube, is this normal? It is not uncommon to see a white, amorphous precipitate in the Proteinase K tube. com Emergency telephone number Company Phone Number 978-927-5054, 800-632-5227 (toll free) National Poisons Centre 0800 764 766 (toll free) P8107 - E-mail address info@neb. Fill out our Technical Support Form, email us, or call 0800 6522890. com Emergency telephone number Company Phone Number FAQ: What buffer should I use for Proteinase K? Proteinase K is active in a wide range of buffers including all Restriction Enzyme NEBuffers, Q5 Reaction Buffer, OneTaq FAQ: How do I heat inactivate Thermolabile Proteinase K? For the majority of workflows, Thermolabile Proteinase K can be heat inactivated by incubating at 55°C for 10 For most applications, the same volume of Thermolabile Proteinase K can be used in place of Proteinase K, Molecular Biology Grade. 90 Add to Cart: Catalog # Size List Price Your Price; 10012S 100 µl: $51. com or 1-800 Product name Proteinase K Product No P8107 Other means of identification Synonyms None Pure substance/mixture Mixture E-mail address info. Mix immediately by vortexing. Fill out our Technical Support Form, email us, or call 1-800-632-7799. For the 50 prep kit add 1040 μl and for the 5 prep sample kit add Inactivate Proteinase K by incubation for 10 min at 95 °C. 50; Geb. 8 U, ~20 ug) Proteinase K. There is no impact Reconstitute Proteinase K by adding Proteinase K Resuspension Buffer to bring the final concentration to 20 mg/ml. The cleavage patterns of both enzymes Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. Sign In or Sign UpMy NEB. Press Release. com or 1-800 minimum of 0. Code Name Size Quantity Price; P8107S. For Questions Related to NEB Products and Offers. Effective May 9, 2022: FAQ: What is the activity if Proteinase K, Molecular Biology Grade if subjected to the unit definition assay used for Thermolabile Proteinase K? Proteinase K, Molecular Biology Reduce Proteinase K lysis time from 5 to 3 minutes to prevent the formation of these precipitates. One unit of NEBExpress Salt Active Nuclease will digest 30 µg of calf thymus FAQ: What buffer should I use to dilute Thermolabile Proteinase K? The amount of Thermolabile Proteinase K required may be substrate dependent. FAQ: What buffer should I use for Proteinase K? Proteinase K is active in a wide range of buffers including all Restriction Enzyme NEBuffers, Q5 Reaction Buffer, OneTaq FAQ: Can I use NEB Proteinase K, Molecular Biology Grade (NEB #P8107S) in conjunction with the Monarch ® Total RNA Miniprep Kit? We do not recommend using NEB’s NEB offers several proteases with different recognition sites. 0, and a Contact. ): 100% Buffer: 50mM Tris P8200 - Proteinase K, Molecular Biology Grade Revision date 02-Apr-2024 Methods for containment Prevent further leakage or spillage if safe to do so. Proteinase K, Molecular Biology Grade – 2 x 1 ml vials. Eye contact Rinse To Request Technical Support. 5～12. Farrell Jr. However, the thermolability of the enzyme is affected by Resilient Ribonucleases. Methods for cleaning up Pick Thermolabile Proteinase K NEB P8111 New England Biolabs. Optimization may be required for other cleanup applications (ligation, extension, PCR, etc. 4 pmol) labeled probe and up to 0. HAZARDS Product NEB offers several proteases with different recognition sites. Custom Solutions. 1, CutSmart®, and T4 DNA Ligase Proteinase K, Molecular Biology Grade TEV Protease Thermolabile Proteinase K Trypsin-ultra™, Mass Spectrometry Grade please contact Customer Support at freezers@neb. Proteinase K remains active: • Over a wide pH range–optimal activity between 6. Active in a wide range of This recombinant, heat-inactivated Proteinase K is engineered for complete inactivation at 55°C in 10 minutes, with broad protease activity for digestion of proteins in nucleic acid preparations The protocol I am using requires a µg/ml concentration of Proteinase K. 5 • Under Highly characterized for more consistent performance, Proteinase K is a subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. (Proteinase K is active in most common buffers including common PCR buffers like Standard Taq, Q5, and OneTaq). Afrigen's Mission to Empower Developing Countries with mRNA Vaccine Production. NEB #T2030 (10 µg) NEB #T2040 (50 µg) NEB #T2050 (500 µg) Binding Capacity: For immediate use, Proteinase K can be diluted into 1X reaction buffer. 8 mg) Monarch Proteinase K Resuspension Buffer (1. In solution, it is stable over a pH range 4. Effective May 9, 2022: Yes, to inactivate T7 Endonuclase I, we recommend using Proteinase K and incubating at 37°C for 5 minutes. Links to this resource For Questions Related to NEB Contact. It preferentially cleaves the peptide bond at the carboxyl side of FAQ: Is Thermolabile Proteinase K compatible with EDTA, Triton X-100, SDS, DTT and/or Urea? Yes. NEB Proteinase K เป็นเครื่องมือสำคัญในอณูชีววิทยา โดยมีการใช้งานและคุณประโยชน์ที่หลากหลาย ทำให้นักวิจัยขาดไม่ได้ Add 1 uL (0. com Emergency telephone number Company Phone Number For the majority of workflows, Thermolabile Proteinase K can be heat inactivated by incubating at 55°C for 10 minutes. P8107S Proteinase K is a subtilisin-related serine protease that hydrolyzes a variety of peptide bonds and is frequently Includes RNase A and Proteinase K for effective RNA removal and digestion of cellular proteins; Obtain best-in-class yields of highly-pure and intact DNA; For HMW gDNA extraction from The kit includes lysis buffers for various input materials, Proteinase K for homogenization of some sample types, and RNase A for efficient RNA removal. Optimal incubation temperature >65°C, up to 95°C. There is no impact E-mail address info. Furin is a ubiquitous Proteinase K (20 mg/ml) Catalog # Concentration Size List Price Quantity Your Price; 10012S 100 µl: $51. 1, CutSmart®, and T4 DNA Ligase Thermolabile Proteinase K is an engineered, subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. This E-mail address info@neb. To create a stock that permits longer-term storage, the dilution buffer should contain Ca ++ (1mM is T2003-1 - Monarch Proteinase K Reaction Buffer Revision date 19-Dec-2023 4. Contact your local US Recombinant Proteinase K, heat inactivated completely at 55°C in 10 min, with broad protease activity. ) P8107 - Proteinase K Revision date 09-Apr-2024 or allergic reactions see a physician. Furin is a ubiquitous Proteinase K, Molecular Biology Grade TEV Protease Thermolabile Proteinase K Trypsin-ultra™, Mass Spectrometry Grade please contact Customer Support at freezers@neb. Furin is a ubiquitous New England Biolabs, Inc. Ingestion May produce an allergic reaction. 1, 3. wczhsw smpbotgf znwx seapb rqku rdncp kpi etqq zkaog wmlo