| Description | Earthworm Fibrinolytic Enzyme (Lumbrokinase) is a type of enzyme extracted and purified from earthworms that can dissolve fibrin. It is present in the digestive tract of earthworms. Earthworm Fibrinolytic Enzyme has a dual function: in addition to directly hydrolyzing fibrin, it can also activate Earthworm Fibrinolytic Enzyme (Lumbrokinase) is a type of enzyme extracted and purified from earthworms that can dissolve fibrin. It is present in the digestive tract of earthworms. Earthworm Fibrinolytic Enzyme has a dual function: in addition to directly hydrolyzing fibrin, it can also activate plasminogen into plasmin, thereby exerting an indirect fibrin-hydrolyzing effect. After animals take Earthworm Fibrinolytic Enzyme, obvious antithrombotic and thrombolytic effects are produced. Lumbokinase attenuates myocardial ischemia-reperfusion (I-R) injury through the activation of Sirt1 signaling, and thus enhances autophagic flux and reduces I-R-induced oxidative damage, inflammation and apoptosis.CharacteristicsThis product is a pale yellow to yellowish-brown powder; it has a fishy odor and is hygroscopic.IdentificationTake an appropriate amount of this product, add water to prepare a solution containing 0.5mg per 1ml, and determine according to the spectrophotometry (Appendix IV A of the 2000 Edition of Pharmacopoeia of the People's Republic of China, Volume II); there shall be a maximum absorption at a wavelength of 278nm.Take an appropriate amount of this product, add 0.9% sodium chloride solution to prepare a solution containing 10mg per 1ml as the test solution; use a No. 6 needle to take 1 drop of animal blood and place it at the bottom of a test tube. After 30 minutes, add 1ml of the test solution to the test tube, shake gently, and the blood clot shall dissolve within 60 minutes. Use 1ml of 0.9% sodium chloride solution as the blank, and perform the same operation; the blood clot shall not dissolve.TestsAcidity and Alkalinity: Take this product, add water to prepare a solution containing 1mg per 1ml, and determine according to the method (Appendix VI H of the 2000 Edition of Pharmacopoeia of the People's Republic of China, Volume II); the pH value shall be 6.0 - 8.0.Clarity and Color of Solution: Take an appropriate amount of this product, add water to prepare a solution containing 1mg per 1ml; the solution shall be clear. If colored, it shall not be darker than the Yellow No. 2 Standard Color Solution (Method 1 of Appendix IX A of the 2000 Edition of Pharmacopoeia of the People's Republic of China, Volume II).Loss on Drying: Take an appropriate amount of this product, use phosphorus pentoxide as a desiccant, dry it under reduced pressure at 60℃ to constant weight; the weight loss shall not exceed 5.0% (Appendix VIII L of the 2000 Edition of Pharmacopoeia of the People's Republic of China, Volume II).Determination of Specific ActivityTiter DeterminationReagents0.01mol phosphate buffer (pH 7.8): Take 3.58g of disodium hydrogen phosphate, add water to dissolve and dilute to 1000ml as Solution A; take 0.78g of sodium dihydrogen phosphate (NaH₂PO₄·2H₂O), add water to dissolve and dilute to 500ml as Solution B; mix Solutions A and B until the pH value reaches 7.8.Working Solution: Mix 0.01mol/L phosphate buffer (pH 7.8) and 0.9% sodium chloride solution at a ratio of 1:17.1.5% Agarose Solution: Take 1.5g of agarose, add 100ml of the working solution, and heat to dissolve.Fibrinogen Solution: Take an appropriate amount of fibrinogen, add the working solution to prepare a coagulable protein solution containing 1.5mg per 1ml.Thrombin Solution: Take thrombin, add 0.9% sodium chloride solution to prepare a solution containing 1 BP unit per 1ml.Preparation of Standard Solution: Take the lumbrokinase standard, add 0.9% sodium chloride solution to prepare solutions with concentrations of 10,000, 8,000, 6,000, 4,000, and 2,000 lumbrokinase units per 1ml respectively.Preparation of Test Solution: Take an appropriate amount of this product, add 0.9% sodium chloride solution to dissolve and dilute to a concentration within the range of the standard curve.Determination Method: Take 39ml of the fibrinogen solution, place it in a beaker, add 39ml of the 55℃ agarose solution and 3.0ml of the thrombin solution while stirring, mix immediately, and quickly pour into a plastic petri dish with a diameter of 14cm. Place it horizontally at room temperature for 1 hour, then punch holes. Precisely measure 10µl each of the lumbrokinase standard solutions of different concentrations and the test solution, spot them on the same petri dish respectively, cover the dish, and place it in a 37℃ incubator for 18 hours of reaction. After taking it out, use a caliper to measure the vertical diameters of the lysis zones. Take the logarithm of the number of units of the kinase standard as the abscissa and the logarithm of the product of the vertical diameters as the ordinate, calculate the regression equation, and substitute the logarithm of the product of the vertical diameters of the test sample into the regression equation to calculate the titer units of the test sample. Both the standard and the test sample shall be tested in duplicate, and the average value shall be used for calculation.Protein ContentTake about 20mg of this product, weigh it accurately, determine according to the nitrogen determination method (Method 2 of Appendix VII D of the 2000 Edition of Pharmacopoeia of the People's Republic of China, Volume II), multiply the result by 6.25 to get the protein content in the test sample, and calculate the milligrams of protein per 1mg of the test sample.Specific ActivityCalculate the specific activity according to the following formula:... Read More | Inquire | Inorganic pyrophosphates are inevitably produced in the process of mRNA transcription in vitro. These substances have a great inhibitory effect on transcription. Inorganic pyrophosphatase (PPase) can hydrolyze the inorganic pyrophosphates produced in nucleic acid amplification experiments, promote Inorganic pyrophosphates are inevitably produced in the process of mRNA transcription in vitro. These substances have a great inhibitory effect on transcription. Inorganic pyrophosphatase (PPase) can hydrolyze the inorganic pyrophosphates produced in nucleic acid amplification experiments, promote the shift of reaction equilibrium to the product generation end, and increase the amount of products.The molecular weight of PPase (pyrophosphatase, inorganic, inorganic pyrophosphatase) is about 63kd, which can catalyze the hydrolysis of inorganic pyrophosphate to produce orthophosphate: P2O74_+H2O+PPase→2HPO42_. In the nucleic acid amplification experiment, PPase can hydrolyze the inorganic pyrophosphate generated with the reaction to avoid its inhibition on the reaction system. The removal of pyrophosphate can shift the reaction equilibrium to the product generation end.This product is a GMP level recombinant inorganic pyrophosphatase (yeast source) expressed by large-scale fermentation of E. coli. It is produced with raw and auxiliary materials of medicinal specifications, and the host protein residue and nucleic acid residue are strictly controlled. The product production and quality management procedures in line with GMP specifications ensure that the production process and all raw and auxiliary materials can be traced.Quality requirements project standard appearance Clear liquid Visible foreign matter Compliance with regulations PH value 7.5±8.5 activity 98U/ml-102U/ml purity ≥95% Endonuclease residues Degradation of substrate shall not exceed 10% Exonuclease residues Degradation of substrate shall not exceed 10% RNase residue Degradation of substrate shall not exceed 10% Bacterial endotoxin content ≤10EU/ml Exogenous DNA residue ≤100pg/mg Host protein residue ≤50ppm Mycoplasma detection negative Heavy metal residues ≤10ppm Follow the following specifications1. ISO 9001:2015, certified facility。2. GMP appendix - cell therapy products State Drug Administration.3. general introduction to human gene therapy - Chinese Pharmacopoeia 2020, National Pharmacopoeia Committee.4. USP chapter <1043>, adjuvant materials for cell, gene, and tissue engineered products.5. USP chapter <92>, growth factors and cytokines used in cell therapy manufacturing.6. Ph. Eur. General chapter 5.2.12, raw materials of biological origin for the production of cell-based and gene therapy medical products.Product features1. hydrolyze inorganic pyrophosphate.2. DNA synthesis: significantly enhance DNA replication ability.3. RNA synthesis: increase RNA production in in vitro transcription reaction.4. The optimal reaction temperature is 25℃, and the enzyme can be inactivated at 65℃ for 10min.Product usage1. optimize RNA transcription: improve the RNA yield of in vitro transcription reaction.2. remove PPI contamination from reagents for SNP genotyping by pyrophosphate assay.3. promote the synthesis of protein, RNA and DNA.4. catalyze the reaction of PPI + H2O → 2pi.5. ssr-pcr optimization：Improve efficiency and increase DNA production.Activity definitionCatalytic inorganic pyrophosphate formation 1 per minute under standard reaction conditions µ The amount of enzyme required for mol phosphate was defined as 1 active unit.Preservation system20 mM Tris-HCl; 100 mM NaCl; 1 mM DTT; 0.1 mM EDTA; 50% (v/v) Glycerol; pH 8.0。 Storage temperature-20±5 ℃。Matters needing attention1. the enzyme has activity in various reaction buffers. Generally, the enzyme can be directly added in HDA, lamp and other experiments.2. the dosage of the enzyme needs to be optimized in different experiments, usually adjusted at the concentration of 0.05~1u/ml.3. the optimum reaction temperature of the enzyme was 25 ℃, and it was active at 16~37 ℃, and the enzyme could be inactivated at 65 ℃ for 10min.4. cofactor: mg2+ is necessary for enzyme activity... Read More | Inquire | Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description： 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hingePurity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description： 100B, previously called S100 beta, belongs to the S100 family within the EF-hand superfamily of Ca2+ binding proteins. S100 proteins contain two EF-hand motifs that differ in affinity, separated by a hinge region with a hydrophobic cleft that is exposed upon Ca2+ binding. S100B is a 91 amino acid (aa) protein, after removal of the initial methionine, and is found as homodimers of 10.4 kDa monomers. Human S100B shares 99%, 98%, 100%, 99% and 97% aa sequence identity with mouse, rat, rabbit, equine and bovine S100B, respectively. Within the S100 family, human S100B shows the highest aa identity (59%) with S100A1. S100B is expressed primarily by astrocytes and oligodendrocytes in the central nervous system, and by Schwann cells in the peripheral nervous system. Ca2+-bound S100B interacts in vitro with at least 20 cytoplasmic proteins, including several structural molecules such as tubulin and GFAP. It can inhibit the phosphorylation of these kinase substrates and others such as tau and neuromodulin. Astrocytes can secrete S100B, which then acts in a cytokine-like manner. Nanomolar concentrations of S100B are secreted constitutively, promote proliferation, and are neurotrophic and anti-apoptotic. Blood levels of S100B reflect extracellular concentrations within the nervous system, and are elevated in Down’s syndrome, Alzheimer’s disease and Tourette’s syndrome, metabolic stress, acute brain injury and brain tumors. Micromolar concentrations of S100B can be destructive and pro-apoptotic; they induce the expression of iNOS, COX-2, IL-1, IL‑6 and TNF-alpha by microglia, astrocytes or neurons. Most extracellular actions of S100B can be mediated by RAGE (receptor for advanced glycation end products), which is also a receptor for other S100 proteins... Read More |