| Description | Aladdin's 10% SDS-PAGE Resolving Gel Master Mix contains almost all reagents required for the preparation of 10% 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 10% SDS-PAGE Resolving Gel Master Mix contains almost all reagents required for the preparation of 10% 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 10% 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 gel10% 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 | Source: Microorganism Isoelectric point: 6.5 Michaelis constant: 9.2×10^-3 M (D-Glucose); 8.6×10^-3 M (NAD) Optimum pH: 9.0~9.5 Fig. 1Optimum temperature: 55℃ Fig. 3pH Stability: 6.0-10.0 (25℃, 24hr) Fig. 2Thermal stability: <50℃ (pH 8.0, Source: Microorganism Isoelectric point: 6.5 Michaelis constant: 9.2×10^-3 M (D-Glucose); 8.6×10^-3 M (NAD) Optimum pH: 9.0~9.5 Fig. 1Optimum temperature: 55℃ Fig. 3pH Stability: 6.0-10.0 (25℃, 24hr) Fig. 2Thermal stability: <50℃ (pH 8.0, 30min) Fig. 4Inhibitors: NEM,SDS Effect of various chemicals: Table 1Reaction:... Read More | ProductsThis product is a high purity genomic DNA extract from 293T cells, agarose gel (0.7%) electrophoresis showed that the size of the DNA extract is more than 15Kb, and basically no degradation, the product is ultimately preserved in TE Buffer, which can be widely used in molecular biology ProductsThis product is a high purity genomic DNA extract from 293T cells, agarose gel (0.7%) electrophoresis showed that the size of the DNA extract is more than 15Kb, and basically no degradation, the product is ultimately preserved in TE Buffer, which can be widely used in molecular biology experiments, such as PCR, enzyme digestion, hybridization, microarray analysis, and other molecular biology experiments.The product was quantified using NanoDrop One at a concentration of 200 ng/µL.Preparation and precautions before useLong-term storage at -20˚C is recommended. Before use, the bottle should be removed from the refrigerator and equilibrated to room temperature and centrifuged before opening the cap for use. Samples should be restored to the sealed state as soon as possible after opening.How to use (take qPCR experiment as an example)1. Amplification template preparationThe samples to be detected were diluted with TE (10 mM Tris-Cl, pH 8.0,1 mM EDTA), and the concentration after dilution was as close as possible to the range of 0.05-10 ng/µL. The samples were placed on ice at 4°C and set aside.2. Standard dilution: according to the following table, firstly dilute Human DNA Standard 1 (100ng/uL) with TE to make 5 different concentrations of standards according to the table below. 10ng/µL of DNA Standard 1 (Std. 1) can be stored stably at -20℃ for 1 month; Std2-5 can only be used on the same day, and should be placed at 4℃ or on ice when not in use for the time being after preparation. When not used temporarily after preparation, it should be stored at 4℃ or on ice.styleCorresponding concentration (ng/µL)Minimum dilution volume (in µL)Std.11010 [100 ng/µL DNA Standard 1] + 90 TEStd.22.520 [Std. 1] +60 TEStd.30.62520 [Std. 2] +60 TEStd.40.1562520 [Std. 3] +60 TEStd.50.039062520 [Std. 4] +60 TE3. qPCR reaction system preparationThe cryopreserved reagents to be used were completely thawed and mixed by inversion several times before preparation, and then briefly centrifuged and prepared for use. 20 µL of the base reaction system was as follows.The base reaction system for 20 µL was as follows:reagents20µL reaction system2×qPCRMix10µLPrimerMixXµLProbeMixXµLTemplate4µLddH2OMake up to 20 µLNote: High Rox model: add 1 µL of 50×High Rox per 50 µL of reaction system; Low Rox model: add 1 µL of 50×High Rox per 500 µL of reaction system.Usually, better results can be obtained with a primer concentration of 0.2 µM, and 0.1-1.0 µM can be used as a reference for setting the range.The concentration of the probe used is related to the fluorescent quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, so please refer to the manual of the instrument or the specific requirements for the use of each fluorescent probe for the adjustment of the concentration during actual use.Prepare a sufficient amount of reaction system mixture as required. After the reaction system has been prepared and mixed thoroughly, add 16 µL per well to the reaction wells. Then add the prepared standard and diluted sample into the corresponding reaction wells, the volume of addition is 4µL/well. TE was added to the blank control tube, and the same amount of TE was added at 4 µL/well.It is recommended to use 20 µL for the reaction, if you need to perform a smaller system reaction, reduce the system components in equal proportion.4. qPCR reaction programThe following is an example of our GoldStar Probe Mixture reaction conditions, which should be improved and optimized according to the PCR product template, primer structure and target fragment size.movetemptimingcirculatepremutability95°C10min1denaturation95°C10sec55Annealing/Extension60°C30sec5Data analysis1. Standard curve productionThe standard curve was plotted with reference to the Excel sheet for data processing. The correlation coefficient R2 of the standard curve should not be lower than 0.98, and the slope should be between -3.1 and -3.6 when the Ct value is the vertical coordinate. If the parameters of the standard curve are unreasonable, it is recommended to repeat the experiment... Read More | Purity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description: Major histocompatibility complex, class II, DR alpha (HLA-DRA) belongs to the MHC class II family. HLA-DRA binds peptides derived from antigens which access the endocytic route of antigen presenting cells (APC) Purity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining. Description: Major histocompatibility complex, class II, DR alpha (HLA-DRA) belongs to the MHC class II family. HLA-DRA binds peptides derived from antigens which access the endocytic route of antigen presenting cells (APC) and presents them on the cell surface for identification by the CD4 T-cells. The peptide binding cleft accommodates peptides of 10-30 residues. The peptides presented by MHC class II molecules are generated mainly by degradation of proteins which access the endocytic route, where they are processed by lysosomal proteases and other hydrolases... Read More | Tyrosine decarboxylase catalyzes the removal of the carboxyl group from tyrosine to produce tyramine and carbon dioxide. Pyridoxal 5'-phosphate is a necessary cofactor. By using the apoenzyme prepared from cells grown on a vitamin B6 deficient medium pyridoxal phosphate may be determined. The Tyrosine decarboxylase catalyzes the removal of the carboxyl group from tyrosine to produce tyramine and carbon dioxide. Pyridoxal 5'-phosphate is a necessary cofactor. By using the apoenzyme prepared from cells grown on a vitamin B6 deficient medium pyridoxal phosphate may be determined. The HOLOenzyme may be used to determine tyrosine, phenylalanine and dihydroxyphenylalanine either manometrically or colorimetrically.L-Tyrosine decarboxylase apoenzyme from Streptococcus faecalis has been used in a study to purify and characterize tyrosine decarboxylase and aromatic-L-amino-acid decarboxylase.L-Tyrosine decarboxylase apoenzyme from Streptococcus faecalis has also been used in a study to investigate the stereospecificity of sodium borohydride reduction of tyrosine decarboxylase.One Unit yields 1µmole of CO2 per minute from L-tyrosine at 37°C, pH 5.5. The APOenzyme activity is measured in the presence of excess pyridoxal phosphate... Read More |