| Description | Products contentProducts IntroductionThe 2.5×TFS Master Mixture is a premixed system for all types of multiplexed PCRs. It contains DNA polymerase, PCR Buffer, dNTPs, Mg2+, stabilizers and enhancers at a concentration of 2.5×, which makes it easy and fast to use.The DNA polymerase Products contentProducts IntroductionThe 2.5×TFS Master Mixture is a premixed system for all types of multiplexed PCRs. It contains DNA polymerase, PCR Buffer, dNTPs, Mg2+, stabilizers and enhancers at a concentration of 2.5×, which makes it easy and fast to use.The DNA polymerase contained in the 2.5×TFS Master Mixture is a genetically engineered recombinant enzyme with 5'→3' DNA polymerase activity and no 5'→3' exonuclease activity; it is a new antibody-modified hot-start enzyme that can effectively reduce non-specific amplification caused by non-specific binding of primer and template or primer dimerization at room temperature. The DNA polymerase is modified by a new type of antibody, which is an antibody-modified hot-start enzyme, and can effectively reduce the non-specific amplification caused by the non-specific binding of primers and templates or primer dimerization at room temperature, and at the same time, it has the excellent features of short activation time, strong amplification ability, high sensitivity, good stability, etc. The unique PCR buffer system and the hot-start enzyme are suitable for the PCR. The unique combination of PCR buffer system and hot starter enzyme significantly improves the PCR amplification efficiency, sensitivity and inhibitor tolerance.The product has a wide range of applications, not only for general and dye-based real-time fluorescence PCR, but also for forensic multiple STR amplification reaction, which can be used in forensic analysis, parentage identification and scientific research and other human genetic identification. caveat1. Before use, please mix the product gently by turning it up and down after it is completely melted and centrifuged briefly.2. Avoid repeated freezing and thawing of the product, which may degrade its performance. This product can be stored at -20℃ for a long time. UsageThe following examples are STR reaction systems and conditions, which should be improved and optimized according to the specific use, template, primer structure, target fragment size and amplification effect.PCR reaction systemExtract the DNA amplification reaction system:Blood Card Direct Expansion Reaction System:Attention:1) When designing the primers, the difference between the Tm of each primer should be minimized, and the difference should be controlled within 5℃ as far as possible. If the amplification efficiency is not high, the concentration of primers can be increased; if non-specific amplification occurs, the concentration of primers can be decreased to optimize the reaction system. For optimal amplification, it is recommended that the primer mixture be vortexed for 10 s and centrifuged briefly before use.(2) The amount of DNA template is usually 0.1 ng-1 ng of human genomic DNA as a reference, and the amount of template input can be adjusted according to the amplification effect to determine the optimal amount of template to use.3) Human genome contamination should be avoided during the operation, and a set of negative control (no DNA) is recommended for the experiment.2. PCR reaction conditionsAttention:1) Two-step PCR reaction program is recommended. If you can not get good results due to low Tm value of the primers or large difference in Tm value between primers, you can try to use three-step PCR amplification, the annealing temperature should be set in the range of 55℃-65℃ as a reference (the annealing temperature is usually 5℃ lower than the Tm value), and the extension temperature should be set at 72℃.(2) When good amplification results are not obtained, the annealing and extension time can be appropriately prolonged to 120 s-150 s. The annealing and extension time can be extended to 120 s-150 s. (3) When the PCR product detection appears to be incomplete plus A, the final extension time can be appropriately extended to 30-40 min. 4) The number of cycles can be set according to the downstream application of the amplified product, if the number of cycles is too small, the amplification is insufficient, the recommended number of cycles is 28-31 cycles.5) Blood card direct amplification can be based on the actual amplification effect to increase the 72 ℃ lysis step to improve the amplification efficiency.6) When using the ABI 9700 Thermal Cycler, perform amplification in MAX mode.7) PCR products can be stored at 2-8°C for short-term storage or at -20°C for long-term storage.Attention:(1) Usually, a primer concentration of 0.2 M can give better results, and 0.1-1.0 µM can be used as a reference for setting the range. If the amplification efficiency is not high, the concentration of primer can be increased; if non-specific reaction occurs, the concentration of primer can be decreased to optimize the reaction system.(2) The final concentration of the probe used is related to the fluorescence quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, please refer to the manual of the instrument or the specific requirements for the use of each fluorescent probe to adjust the concentration.3) Usually the amount of DNA template is 10-100 ng of genomic DNA or 1-10 ng of cDNA as a reference, as templates of different speciesThe number of copies of the target gene contained in them varies, and a gradient dilution of the template can be performed to determine the optimal amount of template to use. PCR reaction conditions :(1) The enzyme used in this product is activated by pre-denaturation at 95°C for 30 s. Most of the templates can be deconvoluted well under this condition. Under this condition, most of the templates can be well unchained. For templates with high GC content and complex secondary structure, the pre-denaturation time can be extended to 1 minute, so that the starting template can be fully unchained, and if the high temperature treatment time is too long, it will affect the activity of the enzyme; for simple templates, pre-denaturation can also be used for 20 s, and the optimal pre-denaturation time can be determined according to the template situation.(2) It is recommended to use two-step PCR program, the annealing temperature should be 58-64℃ as the reference range, and the annealing temperature can be increased in case of non-specific reaction. If you can't get good results due to the use of primers with low Tm value, you can try three-step PCR amplification, and the annealing temperature should be set in the range of 56℃-64℃ as a reference. The annealing and extension times for several common instruments are shown in the following table: 20 s for Roche, BioRad, Agilent, Hongshi, Dongshenglong, etc. 30 s for ABI 7000/7300/7500. The annealing/extension times can be set according to the different types of instruments and templates, please follow the instructions of the instruments. The annealing/extension time can be set according to different models of instruments and templates, please follow the instruction manual of the instrument... Read More | 1、Product attributeReaction time：short (up to 20 minutes) at 20-37°CLot-to-lot variation：<5%Boiling point : 100℃pH-Value (at 20 °C): 3.5-4.0Density (20℃) : 1.0111 g/cm³Appearance: colourless to pale blue liquidOdour: odourlessRecommend Incubation 1、Product attributeReaction time：short (up to 20 minutes) at 20-37°CLot-to-lot variation：<5%Boiling point : 100℃pH-Value (at 20 °C): 3.5-4.0Density (20℃) : 1.0111 g/cm³Appearance: colourless to pale blue liquidOdour: odourlessRecommend Incubation temperature: 20-37 °C2、Requirements for storage rooms and vessels1.Keep container tightly closed.2.Keep cool. protected from light3.Keep/Store only in original container.4.Never return spills in original containers for reuse.5. Keep away from: Food and feeding stuffs3、It is a ready-to-use, labelling-free TMB-substrate solution.4、Biosafety informationThis mixture is not classified as hazardous in accordance with Regulation (EC) No 1272/2008；5、Advantage1. Very high absorbance yield2. Very low background signals3. Certified long-term stability4. Regeneration following light exposure... 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 | Purity>95% SDS-PAGE.FunctionImportant adipokine involved in the control of fat metabolism and insulin sensitivity, with direct anti-diabetic, anti-atherogenic and anti-inflammatory activities. Stimulates AMPK phosphorylation and activation in the liver and the skeletal muscle, enhancing glucose Purity>95% SDS-PAGE.FunctionImportant adipokine involved in the control of fat metabolism and insulin sensitivity, with direct anti-diabetic, anti-atherogenic and anti-inflammatory activities. Stimulates AMPK phosphorylation and activation in the liver and the skeletal muscle, enhancing glucose utilization and fatty-acid combustion. Antagonizes TNF-alpha by negatively regulating its expression in various tissues such as liver and macrophages, and also by counteracting its effects. Inhibits endothelial NF-kappa-B signaling through a cAMP-dependent pathway. May play a role in cell growth, angiogenesis and tissue remodeling by binding and sequestering various growth factors with distinct binding affinities, depending on the type of complex, LMW, MMW or HMW.Post-translationalHydroxylated Lys-33 was not identified in PubMed:16497731, probably due to poor representation of the N-terminal peptide in mass fingerprinting. HMW complexes are more extensively glycosylated than smaller oligomers. Hydroxylation and glycosylation of the lysine residues within the collagene-like domain of adiponectin seem to be critically involved in regulating the formation and/or secretion of HMW complexes and consequently contribute to the insulin-sensitizing activity of adiponectin in hepatocytes. O-glycosylated. Not N-glycosylated. O-linked glycans on hydroxylysines consist of Glc-Gal disaccharides bound to the oxygen atom of post-translationally added hydroxyl groups. Sialylated to varying degrees depending on tissue. Thr-22 appears to be the major site of sialylation. Higher sialylation found in SGBS adipocytes than in HEK fibroblasts. Sialylation is not required neither for heterodimerization nor for secretion. Not sialylated on the glycosylated hydroxylysines. Desialylated forms are rapidly cleared from the circulation... Read More | BackgroundStreptavidin is a tetrameric bacterial protein isolated from Streptomyces avidinii providing 4 high-affinity biotin binding sites. Streptavidin homo-tetramers have an extraordinarily high affinity for biotin. With a dissociation constant on the order of ≈10⁻¹⁴ mol/L,BackgroundStreptavidin is a tetrameric bacterial protein isolated from Streptomyces avidinii providing 4 high-affinity biotin binding sites. Streptavidin homo-tetramers have an extraordinarily high affinity for biotin. With a dissociation constant on the order of ≈10⁻¹⁴ mol/L, the binding of biotin to streptavidin is one of the strongest non-covalent interactions known in nature. Unlike egg-white avidin, which has a net positive charge at neutral pH and contains about 7% carbohydrate, streptavidin has almost no net charge at neutral pH, does not contain carbohydrate, and exhibits lower non-specific background. Streptavidin conjugates are widely used together with a conjugate of biotin for specific detection of a variety of proteins, protein motifs, nucleic acids and other molecules. This FITC-streptavidin conjugate was prepared by highly purified Streptavidin and free FITC was removed. Streptavidin (FITC) is a useful second-step reagent for the indirect immunofluorescent staining of cells in combination with biotinylated primary antibodies for flow cytometric analysis. Excitation at 488nm light leads to a fluorescence emission maximum of 520 nm.Recommended Usage：Every lot of Streptavidin-FITC is tested by flow cytometry using biotinylated primary antibodies. From this testing it is recommended that between 0.02 and 0.25 µg of streptavidin be used per 106 cells in a 100 µl staining volume... Read More |