| Description | Hotstart HiTaq& Script III combines high-speed thermal cycling with exceptional quantification accuracy. Featuring a thermostable reverse transcriptase (HS Script III RTase) and chemical-modified hot-start Taq DNA polymerase (Hotstart HiTaq), it enables contamination-resistant, single-tube Real-Hotstart HiTaq& Script III combines high-speed thermal cycling with exceptional quantification accuracy. Featuring a thermostable reverse transcriptase (HS Script III RTase) and chemical-modified hot-start Taq DNA polymerase (Hotstart HiTaq), it enables contamination-resistant, single-tube Real-Time One-Step RT-PCR with superior sensitivity—ideal for detecting low-abundance RNA targets (e.g., RNA viruses). This master mix is specifically designed for convenient, high-sensitivity one-step RT-PCR. Its unique enzyme system and specially formulated buffer ensure efficient and accurate reverse transcription and PCR amplification without requiring optimization. It delivers excellent standard curves across a broad quantitative range, enabling precise target gene quantification with high reproducibility and reliability. Suitable for hot-start RNA amplification.Product Component ListU1492576Component100T200TStorageU1492576A4×RT PCR Buffer625 µL2.5 mL-20℃U1492576BHotstart HiTaq& Script III100 µL400 µL-20℃.Protocol1. Pre-Reaction SetupThaw components on ice. Avoid freeze-thaw cycles.Prepare reactions on ice.2. Reaction AssemblyComponentVolume/25 µLVolume/50 µLFinal Concentration4×RT PCR Buffer6.25 µL12.5 µL1×Hotstart HiTaq& Script III1 µL2 µL2/50 µLPrimer-Probe Mix1 µL2 µL/Template (RNA)5 µL10 µL/ddH2OTo 25 µLTo 50 µL/Total Volume25 µL50 µL* Enzyme activity: 2 U Hotstart HiTaq + 5 U HS Script III per 50 µL reactionNotes:Adjust primer/probe/template volumes as needed.Protect probes from light.3. Mixing & CentrifugationGently pipette-mix → Centrifuge briefly.4. Cycling ConditionsStepTemperatureTimeCyclesReverse Transcription50℃15min1Initial Denaturation95℃10min1Denaturation95℃10s45Annealing/Extension55℃40s45Hold12℃∞1... Read More | Alanine Aminotransferase (ALT), also known as serum glutamic-pyruvic transaminase (SGPT), is a pyridoxal-phosphate-dependent enzyme that catalyzes the reversible transfer of an amino group from alanine to α-ketoglutarate, generating pyruvate and glutamate. ALT is found primarily in liver and Alanine Aminotransferase (ALT), also known as serum glutamic-pyruvic transaminase (SGPT), is a pyridoxal-phosphate-dependent enzyme that catalyzes the reversible transfer of an amino group from alanine to α-ketoglutarate, generating pyruvate and glutamate. ALT is found primarily in liver and serum, but occurs in other tissues as well. Hepatocellular injury often results in an increase of serum ALT levels and serum ALT levels can be used as a marker for liver injury.ALT Activity Assay kit has been used to determine the activity of alanine aminotransferase (ALT) in serum samples... Read More | Inquire | Inquire | The commonly used method of eukarYOtic gene expression regulation research is the detection of reporter genes, and bioluminescence is the most commonly used and effective means of reporter gene detection. Luciferase can catalyze the conversion of the substrate luciferin and emit photons. This The commonly used method of eukarYOtic gene expression regulation research is the detection of reporter genes, and bioluminescence is the most commonly used and effective means of reporter gene detection. Luciferase can catalyze the conversion of the substrate luciferin and emit photons. This product provides a rapid, sensitive and stable detection method for the expression of Renilla luciferase reporter gene in mammalian cells. Product characteristic：1.Rapid : Cell lysis was completed within 10-15 min ；2.Convenience : The reagent is easy to prepare, and the sample detection steps are simple；Instruction：1. Cell lysis ( 1 ) Remove the culture medium and gently wash with PBS ( adherent cells can be directly performed this operation, suspension cells should be centrifuged to collect cells ). Add 1 × Lysis Buffer ( diluted component A with sterile water at 4 : 1 ) according to the following scheme, and then place the culture plate on a micro-oscillator at room temperature for 15 min to fully lyse the cells. Note : The pyrolysis products can be stored at room temperature for 6 h, and can be stored at − 70 °C for a long time ( the pyrolysis products cannot be repeatedly frozen and thawed ). ( 2 ) The pyrolysis products after full pyrolysis were centrifuged at 10000-15000 rpm for 3-5 min. After centrifugation, the supernatant was moved into a new EP tube for subsequent detection. 2. Preparation of working fluid ( 1 ) Restore all components to room temperature. ( 2 ) Dilute component C into renilla luciferase working solution with component B, and the dilution method is to add 1 µL C component to 49 µL B component. 3.chemiluminescence value detection ( 1 ) According to the operation instructions of the instrument, the instrument with chemiluminescence detection function was opened, such as multifunctional microplate reader. The parameters were set, the determination time was 10 s, and the determination interval was 2 s. ( 2 ) The cell lysis products were added to the measuring tube according to the volume of 20 ~ 100 µL ( keep the same amount of samples each time ). 1 × Lysis Buffer was blank control. ( 3 ) 100 µL renilla luciferase working solution was added to determine the RLU ( Relative light unit ) value ( Shaking mixing function is recommended for microplate reader ). Note : The renilla luciferase working solution cannot be stored for a long time. It is now ready for use and is used once. Component：RenillaLuciferase Lysis Buffer；RenillaLuciferase Assay Buffer；CoelenterazineMatters needing attention：Scope of application： Matters needing attention：1.Please instantaneously centrifuge the product to the bottom of the tube before use, and then carry out subsequent experiments ; 2.Due to the influence of temperature on the enzyme reaction, the sample and reagent should be measured after reaching room temperature. 3.The strongest wavelength of bioluminescence catalyzed by renilla luciferase is 480 nm, in order to prevent interference between holes, it is recommended to use white opaque orifice plate ;4. B component is recommended to carry out small batch packing according to the experimental requirements ; 5.It is recommended to use it now to avoid repeated freezing and thawing ; 6.For your safety and health, please wear experimental clothes and wear disposable gloves. Scope of application：Study on gene expression regulation and promoter... Read More |