Order number: M3116.0050Shipping: shipped at RT, stored at -20°C
Delivery time: 3- 8 working days
For detailed information on the delivery date, please contact Genaxxon.
Genaxxon Proteinase K Low-Temp is a nonspecific endopeptidase derived from an Arctic marine microbial source. It has a broad substrate specificity and is easy to inactivate after use. Starting at a temperature of 60°C, the Genaxxon Proteinase K Low-Temp is heat-inactivated, making it ideal for heat-sensitive samples.
The Proteinase K Low-Temp is most active in the temperature range of 25°-40° C. Recommended inactivation conditions are: Typically up to 15 minutes at 40°C – 65°C (inactivation by denaturation).
It is known that histones and other proteins shield nucleic acids from an optimal interaction with other DNA-binding proteins and enzymes. Genaxxon Proteinase K Low-Temp is ideal for transforming chromatin and other compact nucleic acids into mere DNA. The enzyme is easily inactivated. This allows thermal inactivation at temperatures that provide RNA integrity and avoid the dissociation of dsDNA.
- Easy to deactivate
- Active at high salt content
- Compatible with downstream analyzes
Preparation of chromosomal DNA for PFGE
- Degradation of nucleases (DNase, RNase) in the preparation of nucleic acids, including the recovery of native RNA
- Isolation of genomic DNA from tissue of mouse tails
- Isolation of genomic DNA from cell cultures
Specific activity: >30 U/mg. Foreign activity: RNase and DNase undetectable. Temperature optimum: + 25°C-40°C (pH 7-10). Recommended inactivation conditions: Typically up to 15 minutes at 40°C – 65°C (inactivation by denaturation).
Inhibitors: General Serine protease inhibitors such as PMSF (M3194) >.
Instructions for use: The enzyme is stable for >1 year when stored at -20°C. Genaxxon Proteinase K Low-Temp is stable even when stored at 4°C (>6 months) and room temperature (>4 weeks).
The optimal enzyme activity is between pH 7 and 10. Recommended temperature range for the activity is 25-40°C. The activity of Genaxxon Proteinase K Low-Temp is not dependent on divalent cations such as calcium (Ca2 +) and is therefore active in buffers containing EDTA (<40°C reaction temperature).
Genaxxon Proteinase K Low-Temp is suitable for the digestion of proteins in the presence of SDS (0.2-1%) and urea (1-5 M). The enzyme tolerates 500mM guanidine thiocyanate and 1% Triton X-100 (>50% activity). There is no significant loss of activity at 400 mM NaCl.
Proteinase K is available from Genaxxon as Molbio Grade powder (M3036) > and as solution (20mg / mL, M3037) > or as NGS grade in powder form (M3115) >.
The easy inactivation allows gentler inactivation compared to competing proteases. If being diligent with keeping samples on ice (e.g. RNA samples) when setting up the PCR reactions, it might also be possible to avoid the inactivation step altogether. Suitable for the digestion of proteins in the presence of SDS (0.2-1%) and urea (1-5 M). The enzyme tolerates 500mM guanidine thiocyanate and 1% Triton X-100 (>50% activity). No significant loss of activity can be observed at 400 mM NaCl.
- Source: Recombinantly produced in Pichia pastoris
- Specific activity: >30 Units/mg
- Concentration: 200 U/mL
- Size: 34 kDa
- Specificity: Nonspecific endopeptidase
- Highly active in temperature range of 25–40°C
- pH: 7–10
Typically up to 15 minutes at 40°C – 65°C (inactivation by denaturation).
- Tolerance to typical buffer additives:
- SDS: 0.2–1%
- Urea: 1–5 mM
- Inhibitors: The enzyme is inhibited by general serine protease inhibitors, such as PMSF.
- Storage: The enzyme is stable upon storage at - 20°C for >1 year. Genaxxon Proteinase is also stable upon storage at 4°C (> 6 months) and room temperature (> 4 weeks). Genaxxon Proteinase is stable at dilute concentrations (< 0.1 mg/ml) in solutions.
- Storage buffer:
- 25 mM Tris-HCl, pH 8.0
- 100 mM NaCl
- 50% glycerol
application:Protein digest in DNA samples for example: Typical uses will be direct-PCR settings of single and few cells. Isolation of vaccinia virus DNA. Purification of genomic DNA from bacteria (miniprep).
Unit Definition:One unit is defined as the amount of enzyme that produces one μmol 4-nitroaniline (extinction coefficient 8.8 mM-1 at 410 nm) at 25°C, pH 8.0 per minute (buffer conditions 50 mM Tris-HCl pH 8.0, 1% DMSO, 1 mM Suc-Ala-Ala-Pro-Phe-pNA).
Sicherheits Hinweise / SafetyH-Sätze: H315, H319, H335
GHS-Symbole: GHS07, GHS08
Klassifizierungen / ClassificationEC-Nr: 254-457-8
Dokumente - Protokolle - Downloads
Here you will find information and further literature on Proteinase K Low-Temp - solution (200 units/mL), easy to inactivate. For further documents (certificates with additional lot numbers, safety data sheets in other languages, further product information) please contact Genaxxon biosience at: firstname.lastname@example.org or phone: +49 731 3608 123.
Dokumente - Protokolle - Downloads
This enzyme differs from normal Proteinase K as it is easier to inactivate and gentler on downstream procedures. Its main purpose is to reduce chromatin to naked DNA prior to analysis. It is best fitted to analysis of small amounts of cells, where endogenous proteinases will not have an adverse effect on downstream analysis
Possible protocol for tissue cells:
Seed small amounts of cells in 96-well plate (few thousands?)
Following day – wash and lyse cells in lysis buffer supplemented with AZ Protease (7.5 U/mL)
Transfer 2µL lysate to a 10µL qPCR reaction kept on ice
Run qPCR directly
Heat inactivate might be necessary in some cases (e.g. in case of not working on ice, as this means that used enzymes will be more susceptible for protease activity), then a typical inactivation step might be up to 15 minutes at 40-65°C, depending on degree of inactivation needed.