| Description | 1、Product attributeShelf life: 36 monthsMarker enzyme: Horseradish peroxidase (HRP)Reaction time：short (up to 20 minutes) at 20-25°CLot-to-lot variation：< 10 %Boiling point : 100℃（Calculated flash point）Flash point:：120 °C （1、Product attributeShelf life: 36 monthsMarker enzyme: Horseradish peroxidase (HRP)Reaction time：short (up to 20 minutes) at 20-25°CLot-to-lot variation：< 10 %Boiling point : 100℃（Calculated flash point）Flash point:：120 °C （Calculated flash point）pH-Value (at 20 °C): 4.5-4.9 （Experimental data）Density (20℃) : 1.0064 g/cm³Water solubility: easily solubleAppearance: colourless liquidOdour: odourlessLight sensitiveHeat sensitive2、Requirements for storage rooms and vessels1.Keep container tightly closed.2.Keep cool. Protect from sunlight.3.Keep/Store only in original container.4.Never return spills in original containers for reuse.5. Keep away from: Food and feedingstuffs.6. The solution can be transported at room temperature, but temperatures exceeding 30 °C have to be avoided. Shipping should be completed within one week.7. The solution will still work beyond the expiry date, but a lower sensitivity has to be taken into account and a precipitate may occur.8. Contaminated or leaked out substrate solution from damaged bottles should not be used and has to be destroyed.3、Effective Components and Principle of FunctionA citrate buffer system (pH 4.5 - 5.0) contains the following effective components:Hydrogen peroxide3,3`,5,5`-Tetramethylbenzidine (TMB)precipitating reagentPeroxidase catalyses the decomposition of hydrogen peroxide, taking electrons from TMB (byoxidation). Oxidation of TMB forms a radical cation that stabilizes by dimerisation and shows the typical blue colour. This blue cation is stabilized by forming an insoluble complex with the precipitating reagent. So at positions with horseradish peroxidase activity a blue precipitate will appear. This precipitate is very fine so that even the smallest dots can be coloured regularly.4、Biosafety informationThis mixture is not classified as hazardous in accordance with Regulation (EC) No 1272/2008；Hazardous components：none (according to Regulation (EC) No 1907/2006 (REACH))5、Advantage1. Rapid precipitation2. Strong adherence to plastic surfaces3. Short reaction time4. Intensive dark blue colour5. Homogeneous staining pattern6、Instruction for usageFor bottling consider the following instructions:•Work in a dust free and darkened room.• Pay attention that the solution has no contact with metal parts (leading to catalysis) of yourinstruments. Closed systems of silicon tubes are favoured.• Clean all instruments and vessels very carefully.• Never touch parts of the instruments that are in contact with the solution with the naked hand. Wear powder free gloves.• Close the bottles immediately to minimize the influence of light and dust.• Use bottles that are not permeable to light, made from HDPE or PP.7. General Instructions for the Use on Microchips• This solution should only be used by qualified laboratory staff familiar with the basics of immunological methods.• Note: The substrate solution is very sensitive to impurities. This is why you should never immerse pipette tips in storage bottles. Unused solution is never returned to the bottle! Don't swap the bottles for the cups!• Substrate Solutions Test systems for instrumental measurements on glass or plastic surfaces (polycarbonate or polystyrene).• Depending on the system, it may be sufficient to wash the microarrays after incubation with the conjugate and then cover them with the substrate solution. Arrays can be developed with a rocking motion until visible color appears (approximately 5 to 10 minutes). Measurements can be performed using wet arrays or after washing with distilled water and drying. Dry microchips can be used for recording purposes. (Note: some glues will destroy colored deposits)... Read More | Es Taq DNA Polymerase is an optimized mixed enzyme of Taq and Pfu DNA Polymerase, with 5 '→ 3' DNA polymerase activity, 5 '→ 3' exonuclease activity, and 3 '→ 5' exonuclease activity. Compared with Taq DNA Polymerase, Es Taq DNA Polymerase has excellent performance of high Es Taq DNA Polymerase is an optimized mixed enzyme of Taq and Pfu DNA Polymerase, with 5 '→ 3' DNA polymerase activity, 5 '→ 3' exonuclease activity, and 3 '→ 5' exonuclease activity. Compared with Taq DNA Polymerase, Es Taq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate, and can efficiently amplify DNA fragments. Most of the PCR products amplified with this product contain an "A" base at the 3 'end, which can be directly used for T/A cloning. This product is suitable for conventional PCR reactions and gene cloning reactions that require high fidelity. E665597Component500 UStorageE665597AEs Taq DNA Polymerase, 5 U/µL 100 µL -20℃. Avoid freeze/thaw cycle.E665597B10×PCR Buffer 1.8 mL -20℃. Avoid freeze/thaw cycle.Activity definition:Using activated salmon sperm DNA as a template/primer, the amount of enzyme required to incorporate 10 nmol of deoxyribonucleotide into acidic insoluble substances is defined as 1 active unit (U) at 74 ℃ for 30 minutes.Quality control:After multiple column purifications, SDS-PAGE detected a purity of over 99%; No exogenous nuclease activity detected; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes in the human genome; Store at room temperature for one month without significant changes in activity.1. PCR reaction system Reagent 50 µlReaction system Final concentration 10×PCR Buffer 5 µL 1× dNTP Mix，10 mM each 1 µL 200 µM each Forward Primer，10 µM 2 µL 0.4 µM Reverse Primer，10 µM 2 µl 0.4 µM Template DNA <0.5 µg <0.5 µg/50 µl Es Taq DNA Polymerase，5 U/µl 0.25-0.5 µl 1.25-2.5U/50 µl ddH2O up to 50 µL /Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration µ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system. 2. PCR reaction conditions Step Temperature Time / Pre denaturation 94℃ 2 min / Denaturation 94℃ 30 s 25-35 cycles Anneal 55-65℃ 30 s 25-35 cycles Extend 72℃ 30 s 25-35 cycles Finally extended 72℃ 2 min / Attention:1) In general experiments, if the annealing temperature is 5 ℃ lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of Es Taq DNA Polymerase in this product is 2 kb/min.3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible... Read More | GoldStar Probe Mixture is a premixed system specifically designed for real-time fluorescence quantitative PCR using probe methods (TaqMan, Molecular Beacon, etc.), with a concentration of 2 x, containing GoldStar Taq DNA Polymerase, PCR Buffer, dNTPs, and Mg2+. The operation is simple and convenientGoldStar Probe Mixture is a premixed system specifically designed for real-time fluorescence quantitative PCR using probe methods (TaqMan, Molecular Beacon, etc.), with a concentration of 2 x, containing GoldStar Taq DNA Polymerase, PCR Buffer, dNTPs, and Mg2+. The operation is simple and convenient. Mainly used for detecting genomic DNA target sequences and RNA reverse transcription cDNA target sequences, such as gene expression analysis, copy number analysis, SNP genotype analysis, etc., suitable for fluorescence quantification using different types of probe methods. The GoldStar Taq DNA Polymerase contained in this product is a chemically modified, novel and highly efficient hot start enzyme. It has no polymerase activity at room temperature, effectively avoiding non-specific amplification caused by non-specific binding of primers and templates or primer dimerization at room temperature. The enzyme activation requires incubation at 95 ℃ for 10 minutes. The unique combination of PCR buffer system and hot start enzyme significantly improves the amplification efficiency of PCR, with stronger fluorescence signal and higher sensitivity, which can detect single copy templates. By using this product, a wider linear range can be obtained, resulting in more accurate quantification of the target gene. Suitable for all fluorescence quantitative PCR instruments that do not require ROX as a calibration dye.ROX dye is used to correct the fluorescence signal error between wells in quantitative PCR instruments, and is generally used in Real Time PCR amplification instruments from companies such as ABI and Stratagene. The excitation optical systems of different instruments vary, so the concentration of ROX dye must be matched with the corresponding fluorescence quantitative PCR instrument.Instruments that do not require ROX calibration (CW0932): Roche LightCycle 480, Roche LightCyler 96, Bio rad iCyler iQ, iQ5, CFX96, etc.Instrument requiring Low ROX calibration (CW2625): ABI Prism7500/7500 Fast, QuantStudio ® 3 System, QuantStudio ® 5 System, QuantStudio ® 6 Flex System, QuantStudio ® 7 Flex System, ViiA 7 System, Stratagene Mx3000/Mx3005P, Corbett Rotor Gene 3000, etc.Instruments that require High ROX calibration (CW2626): ABI Prism7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus, etc.G665832Component5 mLStorageG665832A2×GoldStar Probe Mixture5×1 mL-20℃. Avoid freeze/thaw cycle.G665832BddH2O5×1 mL -20℃. Avoid freeze/thaw cycle. Notes:1. Before use, please gently mix upside down to avoid foaming, and use after briefly centrifugation.2. Avoid repeated freezing and thawing of this product, as repeated freezing and thawing may cause a decrease in product performance. This product can be stored for a long time at -20 ℃, away from light. If frequent use is required in the short term, it can be stored at 2-8 ℃.Usage:The following are examples of conventional PCR reaction systems and reaction conditions. In practical operation, corresponding improvements and optimizations should be made based on different templates, primer structures, and target fragment sizes.1. PCR reaction system Reagent 50 µl Reaction system Final concentration 2×GoldStar Probe Mixture 25 µl 1 × Forward Primer，10 µM 1 µl 0.2 µM¹⁾ Reverse Primer，10 µM 1 µl 0.2 µM¹⁾ Probe，10 µM 1 µl 0.2 µM²⁾ Template DNA 2 µl³⁾ / 50×Low ROX or High ROX（optional）⁴⁾ 1 µl 1 × ddH2O up to 50 µl / Attention:1) Typically, the primer concentration is 0.2 µ M can achieve good results, ranging from 0.1 to 1.0 µ M serves as a reference for setting the range.2) 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. Please refer to the instrument manual or the specific usage requirements of each fluorescent probe for concentration adjustment during actual use.3) The amount of DNA template is usually based on 10-100 ng genomic DNA or 1-10 ng cDNA as a reference. Due to the different copy numbers of target genes contained in templates of different species, gradient dilution can be applied to the template to determine the optimal template usage.4) The excitation optical systems of different instruments vary, and depending on the instrument used for fluorescence quantification, 50 x Low ROX or 50 x High ROX can be added.2. PCR reaction programAttention! The pre denaturation reaction of this product must be completed at 95 ℃ for 10 minutes!Two step PCR Step Temperature Time / Pre denaturation 95℃ 10 min¹⁾ / Denaturation 95℃ 15 s 35-40 cycles Annealing/Extension ²⁾ 60℃ 1 min 35-40 cycles Attention:1) The hot start enzyme used in this product must be activated under pre denaturation conditions of 95 ℃ and 10 minutes.2) It is recommended to use a two-step PCR reaction program. If good experimental results cannot be obtained due to the use of primers with lower Tm values, a three-step PCR amplification can be attempted. The annealing temperature should be set within the range of 56 ℃ -64 ℃ as a reference... Read More | Inquire | H-7 dihydrochloride blocks human immunodeficiency virus (HIV-1) replication in MOLT-4 (clone No. 8) cell line. It increases the secretion of interleukin 1β (IL-1β).Application：H-7 dihydrochloride has been used to study H-7-induced inhibition of contractility in rat embryo H-7 dihydrochloride blocks human immunodeficiency virus (HIV-1) replication in MOLT-4 (clone No. 8) cell line. It increases the secretion of interleukin 1β (IL-1β).Application：H-7 dihydrochloride has been used to study H-7-induced inhibition of contractility in rat embryo fibroblasts (REF52) cells and acts as a kinase inhibitor... Read More |