23 results about "Protease K" patented technology

The invention relates to the field of cosmetic compositions, in particular to a cosmetic composition containing triple polishing enzyme and a preparation method of the cosmetic composition. The triple polishing enzyme comprises protease, protease K and superoxide dismutase, the weight part ratio of the protease to the protease K to the superoxide dismutase is 1: (0.5-2): (0.5-2), the total mass percent of the triple polishing enzyme in the composition is 0.0002%-0.001%, and the invention further discloses a preparation method of the cosmetic composition. A triple polishing enzyme synergistic system is formed through scientific compounding, the integrated skin care effect under low concentration is achieved, and the defects of a traditional chemical stripping agent and the shortages of a single enzyme preparation are overcome; meanwhile, by optimizing the proportion of formula components, the composition forms a liquid crystal structure, so that long-acting dual moisturizing of internal water locking and external water sealing is realized, triple polishing enzyme can be wrapped and protected, and the problems that the composition is easy to oxidize and inactivate and low in transdermal efficiency are solved.

The invention relates to the field of DNA extraction of leather cultural relics, and discloses a method for extracting cattle genomes from vegetable tanned cattle leather cultural relics. The method comprises the following steps: after cleaning the vegetable tanned cow leather cultural relics, dehydrating, freeze-drying and grinding and crushing with liquid nitrogen; metal ions are chelated through an EDTA-Tris detanning reagent to destroy a tannin structure, so that the wrapping on DNA (Deoxyribose Nucleic Acid) is reduced; then, lysate and protease K are used for lysis and digestion, cell membranes are damaged, protein is degraded, and DNA is released; adding absolute ethyl alcohol to precipitate and separate out DNA; and finally, purifying, recovering and purifying the DNA through an adsorption column. The genome DNA extracted by the method is large in fragment, high in purity and stable and reliable in quality.

The invention belongs to the technical field of food detection, and particularly relates to a detection method for determining the form of trace selenium in vegetables in a geological high background area. The method comprises the following steps: cleaning a vegetable sample, freeze-drying, grinding and crushing, adding a specific compound enzyme system (protease XIV, protease K and cellulase) and a Tris-HCl buffer solution, performing water bath ultrasonic treatment, performing constant-temperature oscillation for enzymolysis, performing low-temperature centrifugation on an extracting solution for multiple times, and filtering and purifying the extracting solution by a microporous filter membrane, the method comprises the following steps: detecting the selenium form of a sample by adopting a high performance liquid chromatograph and inductively coupled plasma mass spectrometer serial device, and quantitatively analyzing tetravalent selenium (Se4 +), hexavalent selenium (Se6 +), selenomethionine (SeMet), methylselenocysteine (SeMeCys) and selenocystine (SeCys2) in the sample by adopting a normalization method. The detection method disclosed by the invention is strong in anti-interference capability and high in sensitivity, and original form information can be completely reserved.

The invention relates to an immunofluorescence method and kit for detecting zebra fish heart DNA methylation level, and belongs to the technical field of biological detection. The immunofluorescence method provided by the invention comprises the following steps: exposing a zebrafish embryo to a substance to be detected, dissecting a heart sample of the zebrafish embryo and carrying out fixation, membrane permeation, protease K digestion, hydrochloric acid DNA denaturation and neutralization treatment, and then carrying out sealing, incubation of a 5-methylcytosine primary antibody and a fluorescently labeled secondary antibody, cell nucleus staining and fluorescence detection. And finally, analyzing and evaluating the DNA methylation level through fluorescence imaging. According to the method disclosed by the invention, through a combined step of protease K and hydrochloric acid denaturation treatment, the binding efficiency to the antibody is remarkably improved, so that high-sensitivity and low-background detection on the methylation level of the whole DNA in a zebra fish micro heart sample is realized; the detection kit provided by the invention can be widely applied to epigenetics evaluation of toxicity of environmental pollutants.

The application discloses a DNA / RNA reagent kit based on a magnetic bead method and free of enzyme digestion, and belongs to the technical field of nucleic acid extraction and purification. The reagent kit comprises a lysis solution AL, protease K, a specific binding buffer CLB, amino magnetic bead suspension, silicon hydroxyl magnetic bead suspension and other components. The application solves the problems of nucleic acid degradation, incomplete removal of cross contamination and complicated process caused by enzyme digestion in the prior art, is suitable for automatic high-throughput processing, and has a wide application prospect in scientific research and clinical application.

The invention belongs to the technical field of molecular detection, and discloses a nucleic acid hybridization capture immunofluorescence detection kit as well as a use method and application thereof. In the kit provided by the invention, guanidine hydrochloride, guanidine isothiocyanate, triton X-100, NaCl, protease K and lauryl sodium sulfate are compounded according to a specific concentration to obtain the nucleic acid release liquid, so that pathogen nucleic acid in a clinical sample can be fully dissociated, and the nucleic acid release liquid can cooperate with a probe aiming at a specific target pathogen; the solution system can be used for optimizing the recognition and combination process of DNA and RNA, the specific recognition and combination capacity between the probe and specific pathogen nucleotide is improved, and under the condition that a common nucleic acid stabilizer, namely Tween, is not contained, the solution system can further strengthen differential structural epitopes existing in a DNA / RNA complex compared with DNA and RNA, so that the specificity of the DNA / RNA complex is improved, and the specificity of the DNA / RNA complex is improved. The kit can be better recognized and combined by an anti-DNA / RNA complex antibody, and has excellent detection specificity, sensitivity and precision.

The application discloses a marine organism muscle tissue DNA extraction kit and application thereof, and belongs to the technical field of biological detection. The extraction kit comprises a lysis solution, a first washing solution and a second washing solution. The lysis solution is composed of Triton x-100 45 mM, Tris-HCl 10 mM, EDTA-Na 80 mM, NaCl 80 mM and Brij-35 3 mM, and has a pH of 8.3. The first washing solution is composed of 70% ethanol solution containing 200 mM Tris-HCl. The second washing solution is composed of 80% ethanol solution. The marine organisms include oceanic fishes, Antarctic fishes, shellfishes, crabs and shrimps. The tissue is taken, cut, mixed with the lysis solution and protease K, and uniformly mixed in a water bath. The application is based on the phenol extraction method, and various mild reagents are mixed to realize rapid and accurate DNA extraction, so that the purity, quality and integrity of the DNA are ensured.

The invention discloses a kit capable of being used for extracting specific short fragment cfDNA in urine and application of the kit. The kit comprises protease K, a lysis solution, a cleaning solution, an eluent and magnetic beads, wherein the magnetic beads comprise silicon hydroxyl magnetic microspheres and magnetic microspheres modified by streptavidin. According to the technical scheme, through matching of composition control of the magnetic beads and optimization of selection of the components, when the obtained kit is used for extracting free DNA in urine, the extraction efficiency of specific short fragments is greatly improved, and the detection accuracy is further improved.

The invention belongs to the technical field of medical detection, and discloses a staining reagent applied to acid-fast staining and a staining method. The dyeing reagent comprises a primary dyeing agent, a decolorizing agent, a counterdyeing agent and an accelerating agent, wherein the accelerating agent is prepared from the following components: lauryl sodium sulfate, triton X-100, hexadecyl trimethyl ammonium bromide, a Tris-HCl buffer solution, ethylenediamine tetraacetic acid, sodium chloride, potassium chloride, protease K and beta-mercaptoethanol. According to the dyeing reagent, the dyeing accelerant is introduced into a traditional acid-fast dyeing system, the component capable of specifically damaging the lipid structure of the cell wall of the Mycobacterium is utilized, the hindering effect of the lipid structure on the dye in the dyeing process is reduced, the binding efficiency of the dye and thalli is remarkably improved, and therefore the positive detection rate of acid-fast dyeing is greatly increased.

The invention discloses a targeted protein degradation robot based on DNA origami as well as a construction method and application of the targeted protein degradation robot, and relates to the technical field of biological medicines. The targeted protein degradation robot comprises a DNA origami structure and a recognition module, the construction method of the DNA origami structure comprises the following steps: mixing M13 ssDNA with 60 stapled chains, carrying out annealing reaction to obtain DNA six-screw column origami, and carrying out incubation reaction on the DNA six-screw column origami and a P chain coupled with protease K to obtain origami coupled with inner layer protease K; performing incubation reaction on the origami coupled with the inner layer protease K, a T + S + A-ABD compound and an L + A-ABD compound to obtain an origami compound; and carrying out incubation reaction on the origami compound and human serum albumin (HSA) to obtain the DNA origami structure. The robot can achieve non-endogenous-dependence in-situ precise degradation of target protein, and a new strategy is provided for achieving treatment of diseases such as tumors through interference of the protein level.

The application discloses a preparation method and application of PDRN (plant-derived RNA) from rice callus. The preparation method comprises the following steps: inducing and suspending culturing of the rice callus; homogenizing the rice callus suspension cells obtained in step 1; sequentially using cellulase, pectinase and protease K for enzymolysis; nucleic acid purification and precise ultrasonic shearing are performed in sequence, so that PDRN with a molecular weight of 25-500 bp is obtained. Through optimization of the rice callus culture condition and combination of the joint lysis process of "mechanical grinding-composite enzymolysis-protease treatment", efficient and high-purity preparation of the plant source PDRN is realized, the extraction rate can be higher than 0.5 mg / g of fresh weight of the callus, and the purity A260 / A280 is greater than or equal to 1.8. The obtained PDRN has significant effects in terms of anti-wrinkle, soothing, moisturizing, anti-inflammatory and anti-aging.

The application discloses a crab meat DNA extraction kit and an extraction method, and belongs to the field of molecular biology. The kit comprises a lysis solution, and the lysis solution comprises SDS, protease K and RNase A. The kit is simple in components, easy to prepare, low in cost and free of toxicity. The extraction method is simple and easy to operate, and crab meat DNA can be quickly extracted, thereby promoting the high-quality and sustainable development of the aquaculture industry.

The invention relates to a pretreatment reagent and method for a tissue sample and microorganisms difficult to break walls and application. The pretreatment reagent comprises a first enzyme mixed solution and a second enzyme mixed solution, the first enzyme mixed solution contains protease K and collagenase I, and the second enzyme mixed solution contains lywallzyme and beta-glucanase. A specific pretreatment reagent and a treatment method are designed, a tissue sample is efficiently dissociated into a dispersed cell state by utilizing a first enzyme mixed solution, subsequent grinding and second enzyme mixed solution treatment are facilitated, and the second enzyme mixed solution can greatly improve the release effect of nucleic acid in the sample and has a relatively great promotion effect on subsequent nucleic acid extraction; in addition, specific grinding condition parameters are designed, nucleic acid release can be further promoted, the sample detection performance can be improved, and the probability of false negative in detection is reduced.

The invention relates to the technical field of biomedicine, in particular to a trace blood DNA (deoxyribonucleic acid) extraction method which comprises the following steps: S1, mixing a blood sample and lysate according to a ratio of 1: 3, uniformly mixing by vortex, and centrifuging; s2, protease K is added, vortex mixing is carried out, incubation is carried out at 55 DEG C for 15 min, centrifugation is carried out for 30 s, and 500 microliters of a binding solution is added; and S3, completely transferring the liquid into a purification column with a collection tube, and standing for 2 minutes. The extraction method of the trace blood DNA comprises the following steps: denaturating protein through SDS (Sodium Dodecyl Sulfate), and separating nucleic acid; the binding liquid adopts high salt and low PH, so that the long fragment nucleic acid can be more easily bound on the adsorption column, guanidine hydrochloride can enhance the splitting effect, and the nucleic acid in the blood sample can be fully released due to the dual splitting effect; and on the other hand, the key point is that the step of removing red blood cells by red blood cell lysate is avoided, but the whole blood is directly lysed, so that the loss of white blood cells is reduced, the operation steps are simplified, and the method is suitable for extracting fresh and frozen anticoagulant blood samples.

The invention discloses a method for extracting giardia and cryptosporidium nucleic acid and application thereof. The method comprises the following steps: mixing a sample containing giardia and / or cryptosporidium with lysozyme and protease K, and incubating and digesting; adding a lysis enhancement solution and glass beads into the digested sample, carrying out ultrasonic treatment, and centrifuging after ultrasonic treatment to obtain a supernatant; and carrying out automatic nucleic acid extraction and purification on the obtained supernate. According to the method, the sample treatment time is shortened to be within 3 h, the detection efficiency is greatly improved, the detection cost is reduced, the cracking effect is excellent, the sensitivity is high, and the specificity is high.

The invention relates to a method for quantitatively detecting mycoplasma nucleic acid copy number based on a microdroplet dPCR technology and application, and the method comprises the following steps: adding a lysate into a mycoplasma strain suspension to be detected for lysis, adding protease K for incubation to obtain DNA, diluting, and carrying out dPCR detection; the lysate is prepared from SDS (Sodium Dodecyl Sulfate), Tween-20, Triton X100, EDTA (Ethylene Diamine Tetraacetic Acid), Tris-HCl, beta-mercaptoethanol and sodium chloride; the mycoplasma strain is Mycoplasma hyorhinis, and the mycoplasma strain is Mycoplasma hyorhinis. According to the invention, two cracking reagents are used for cracking a strain suspension, and the cracking solution is diluted by at least 100 times with molecular biological water and then is used for dPCR detection, so that the interference of impurities on an amplification system is reduced; meanwhile, a nucleic acid extraction step is omitted, so that the loss in the nucleic acid extraction process is effectively avoided. Due to the fact that a large amount of free nucleic acid exists in supernate, the method adopts a strain suspension direct cracking mode, and detection and analysis of two parts of nucleic acid of sediment and the supernate can be achieved at the same time.

The invention discloses a multifunctional biological scaffold as well as a preparation method and application thereof. The method comprises the following steps: performing proteinase K treatment on a natural egg membrane to obtain a pure substrate; performing gas foaming in the wet substrate by hydrolyzing sodium borohydride to release hydrogen so as to construct a three-dimensional porous scaffold; then, a polydopamine coating is formed on the surface of the stent through dopamine oxidation auto-polymerization, and nano-silver particles are loaded through in-situ reduction; and finally, through simulated body fluid mineralization, a bone-like apatite layer is deposited on the surface of the coating. A uniformly interconnected three-dimensional porous structure is created through gas foaming, and a physical platform is provided for subsequent functionalization. The obtained final material integrates a three-dimensional porous structure, rapid physical-chemical synergistic hemostasis, lasting broad-spectrum antibacterial property and strong osteogenic activity promotion, and is especially suitable for repairing infectious bone defects or refractory soft tissue wounds. The preparation technology is simple, raw materials are easy to obtain, and high-value utilization of waste resources is achieved.

The invention relates to the technical field of molecular biology and in-vitro diagnosis, in particular to a kit and method for directly extracting circulating free DNA (cfDNA) from whole blood, the kit comprises a stationary liquid, a binding liquid, protease K, magnetic beads, a washing liquid, a rinsing liquid A, a rinsing liquid B and an eluent, and the stationary liquid comprises 20% methanol, 0.8 M trehalose and the balance purified water. According to the kit and the method for directly extracting the circulating free DNA (cfDNA) from the whole blood, the process is simplified, plasma separation is not needed, and the manual processing time and the manual error are reduced; cfDNA loss is reduced, and degradation in the plasma transfer process is avoided; the device is suitable for being integrated to a full-automatic nucleic acid extraction platform; the method is high in extraction efficiency, suitable for low-abundance mutation detection and high in sensitivity; and consumables and manual operation cost are reduced.

The present application relates to the technical field of extraction reagent, in particular to an economic and general high molecular weight DNA extraction reagent and extraction method, which comprises a lysis solution, a precipitant, a binding solution, a washing solution 1, a washing solution 2, an eluent and a protease K. The present application adopts a precipitation method to extract DNA, avoids using high-priced nucleic acid capture carriers (such as magnetic beads and silica gel membranes) and special extraction instruments, and the reagent components are all conventional inexpensive reagents, so that the cost is reduced by more than 30% compared with existing commercial reagents, and the present application is easy to popularize and widely apply in laboratories.

The invention discloses a method for rapidly detecting yeast pollution of fruit granule milk. The method comprises the following steps: S1, treating a fruit granule milk sample by adopting a normal saline pretreatment solution containing protease K, EDTA (Ethylene Diamine Tetraacetic Acid) and polyethylene glycol 8000; s2, filtering and centrifugally removing impurities from the sample treated in the step S1; and S3, taking the total DNA of the sample, and detecting the yeast in the fruit grain milk by adopting a fluorescent quantitative PCR (Polymerase Chain Reaction) method. According to the method, the sterile normal saline containing protease K, EDTA and polyethylene glycol 8000 is adopted to pretreat the fruit grain milk sample, and then a double impurity removal method is adopted to remove impurity components in the fruit grain milk, so that physical interferents are effectively removed, target bacteria are concentrated, the DNA extraction purity is improved, and meanwhile, the detection rate of a low-pollution-degree sample is also improved. Moreover, the detection time is greatly shortened, the whole process from the sample to the result can be completed within 4 hours, which is far better than the traditional process of 3-5 days, so that the real'rapid detection 'is realized, and the detection sensitivity and accuracy are remarkably improved.

The invention discloses a recombinant III-type humanized collagen enzyme digestion peptide-oxidized sodium alginate complex as well as a preparation method and application thereof. The complex is formed by crosslinking and compounding recombinant III-type humanized collagen subjected to enzyme digestion treatment by specific protease and oxidized modified sodium alginate; wherein the specific protease is selected from one or more of Lys-C, Glu-C, protease K, pepsin or trypsin. The recombinant III-type humanized collagen is subjected to enzyme digestion treatment to form a specific polypeptide fragment which can be crosslinked with the oxidized and modified sodium alginate to form a complex which takes a modified sodium alginate polysaccharide molecular chain as a main chain and takes recombinant III-type humanized collagen enzyme digestion polypeptide as an auxiliary chain; therefore, the adhesion effect of the recombinant III-type humanized collagen in the skin is improved, the acting time of the recombinant III-type humanized collagen on the surface of the skin is prolonged, the utilization of the cell level is facilitated, and the bioremediation capability and the moisturizing performance of the recombinant III-type humanized collagen are improved.

The invention discloses an anticoagulant peptide derived from deep sea shellfish and an application of the anticoagulant peptide in antithrombosis. Through separation, purification and structural identification, three anticoagulant polypeptides are obtained from pepsase zymolyte of deep-sea Gigandas plasmos, and the amino acid sequences of the three anticoagulant polypeptides are respectively TVTRLSD, VSQPRSTL and VVTRVVR. The anticoagulant polypeptides have the advantages that the anticoagulant polypeptides are prepared from the pepsase zymolyte of the deep-sea Gigandas plasmos, and the anticoagulant polypeptides are prepared from the pepsase zymolyte of the deep-sea Gigandas plasmos; the method comprises the following steps: obtaining polypeptide KDETRTPEEL from protease K zymolyte of deep-sea metaphila palustris; the polypeptide YLERW is obtained from the protease K zymolyte of the deep sea hippocampus (Gigantidas hamaensis), and the polypeptide YLERW is obtained from the protease K zymolyte of the deep sea hippocampus (Gigantidas hamaensis). An in-vitro anticoagulation experiment result shows that the five deep sea polypeptides can remarkably prolong activated partial thromboplastin time (APTT) and show a remarkable anticoagulation effect. Wherein the inhibition activity on the blood coagulation factor Xa and thrombin is not obvious, and good selective inhibition characteristics are shown. The novel anticoagulant peptide with blood coagulation factor XIa selective inhibitory activity is separated and identified from deep sea shellfish resources, and a novel material basis and a technical approach are provided for prevention and treatment of thrombotic diseases and development of novel biological anticoagulant preparations.

The invention discloses an extraction method of trace plasma cfDNA, a reagent composition and a kit. The method comprises the following steps: centrifugally separating plasma from whole blood; performing heat treatment on the plasma to denature protein; adding a nucleic acid separating medium containing thermosensitive protease K for enzymolysis, and digesting histone combined with the cfDNA; adding a nucleic acid copolymerization solution containing linear acrylamide with a specific concentration and polyethylene glycol 8000, and enriching cfDNA through a molecular crowding effect; finally, purified magnetic beads are used for elution, and high-purity cfDNA is obtained. The invention also provides a nucleic acid separation liquid, a nucleic acid copolymerization liquid and a kit special for the method. The method is simple, convenient and rapid to operate, has high recovery rate and good repeatability on trace plasma, can effectively avoid oxidative damage, and completely retains original methylation information of cfDNA.