| Description | Aladdin's 12% SDS-PAGE Resolving Gel Master Mix contains almost all reagents required for the preparation of 12% SDS-PAGE resolving gels, except for polymerization catalysts such as ammonium persulfate and TEMED.Aladdin's SDS-PAGE Resolving Gel Master Mix series can be used to prepare 5 common Aladdin's 12% SDS-PAGE Resolving Gel Master Mix contains almost all reagents required for the preparation of 12% SDS-PAGE resolving gels, except for polymerization catalysts such as ammonium persulfate and TEMED.Aladdin's SDS-PAGE Resolving Gel Master Mix series can be used to prepare 5 common concentrations of gelsPrecautions：Ammonium Persulfate or its substitute (, ST005) and TEMED are required but not provided in this product.This product contains Acr-Bis which is potentially neurotoxic. Please take effective measures to avoid direct contact with the human body or inhalation.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1.According to the size of a target protein, decide an appropriate concentration of SDS-PAGE resolving gel. Please refer to the table below.SDS-PAGE Resolving Gel Concentration Optimal Separation Range6%50-150kD8%30-90kD10 -80kD12%12-60kD15%10-40kD2.Prepare a 10% ammonium persulfate solution with ddH2O or other high-purity water. Solution of ammonium persulfate or its substitute is prone to failure and should be prepared freshly or kept frozen for multiple uses.3.Prepare the resolving gel according to the table below. For example, add 100µl of 10% ammonium persulfate and 4µl of TEMED into 10ml of 12% SDS-PAGE Resolving Gel Master Mix, mix well, and immediately pour into gel cassette. Overlay with the top layer of water-saturated butanol, isopropanol, 0.1% SDS or distilled water. Leave at room temperature (~25℃) until fully solidified (usually within 10-30 minutes). Note: The amount of polymerization catalyst in the table below is recommended for polymerization at 25℃. It can be adjusted appropriately according to the temperature. For example, when the room temperature is lower than 20℃, add more TEMED and 10% ammonium persulfate appropriately to promote gel solidification.ReagentsVolume of each component (ml) required for different volumes of SDS-PAGE resolving gel12% SDS-PAGE Resolving Gel Master Mix5101520305010% ammonium persulfate or its substitute0.050.10.150.20.30.5TEMED 0.0020.0040.0060.0080.0120.024.After complete polymerization of the resolving gel, remove the liquid used for sealing, and then prepare the SDS-PAGE stacking gel with 's SDS-PAGE Stacking Gel Master Mix. 5.If the prepared gel is not to be used on the same day, it can be stored at 4℃ for 1-2 days... Read More | Inquire | Gly-Pro-pNA hydrochloride is a dipeptidyl peptidase inhibitor that inhibits dipeptidyl peptidase II, dipeptidyl peptidase IV and dipeptidyl peptidase IX | 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 |