| Description | This product is a reagent kit for reverse transcription of genomic DNA, which can remove genomic DNA in 2 minutes at 42 ℃. Meanwhile, the reverse transcription reagent contains components that inhibit gDNA Remover, and the samples treated with gDNA Remover can be directly subjected to reverse This product is a reagent kit for reverse transcription of genomic DNA, which can remove genomic DNA in 2 minutes at 42 ℃. Meanwhile, the reverse transcription reagent contains components that inhibit gDNA Remover, and the samples treated with gDNA Remover can be directly subjected to reverse transcription reaction to synthesize cDNA.This reagent kit is equipped with a novel and efficient reverse transcriptase HiFiScript, which significantly enhances the transcriptional activity of the enzyme through novel mutation sites. Meanwhile, reverse transcription reaction only takes 15 minutes to complete the synthesis of the first strand of cDNA. 5 × HifiScript RT MasterMix is a reverse transcription premix that contains all the reagents required for reverse transcription, making it easy and fast to operate.H665909Component100 TStorageH665909A10×gDNA Remover Mix 100 µL-20℃. Avoid freeze/thaw cycle.H665909B5×HiFiScript RT MasterMix 400 µL-20℃. Avoid freeze/thaw cycle.H665909CRNase-Free Water 1.5 mL-20℃. Avoid freeze/thaw cycle. Product features1. Quick genome removal: gDNA Remover containing genomic DNA can remove genomic DNA in just 2 minutes.2. Rapid reverse transcription: cDNA first strand synthesis can be completed in 15 minutes.3. Convenient and fast: Ready to use reverse transcription Mix, easy to operate.4. High sensitivity: cDNA first strand can be synthesized using pg level total RNA or mRNA templates.5. Efficient reverse transcription efficiency: Novel mutation sites significantly enhance enzyme activity performance, resulting in higher yields of cDNA.Matters needing attention1. During the operation, RNase contamination should be avoided to prevent RNA degradation or cross contamination during experiments. It is recommended that operators wear masks and disposable gloves, frequently change gloves, and use specialized instruments and consumables.2. Disposable plastic containers should be used as much as possible for experiments. If glassware is used, a 0.1% DEPC (diethyl carbonate 1. ester) aqueous solution should be treated at 37 ℃ for 12 hours, and then sterilized under high pressure at 120 ℃ for 30 minutes before use. Alternatively, glassware should be sterilized under dry heat at 180 ℃ for 60 minutes before use. The sterile water used in the experiment should be treated with 0.1% DEPC and then subjected to high-pressure sterilization.3. The reverse transcription system is prepared on ice for operation to prevent RNA degradation. The enzyme in the reagent kit should be stored at -20 º C as soon as possible after use, and repeated freeze-thaw should be avoided as much as possible.UsageThaw the template RNA on ice; After thawing at room temperature, immediately place the components of the reagent kit on ice. Before use, mix each solution by vortex oscillation and centrifuge briefly before use.1、 Genomic DNA removal reaction1. Prepare a reaction system on ice according to the following table, with a total volume of 10 µ L. To ensure the accuracy of the preparation of the reaction solution, a pre mixed system is first prepared in the amount of reaction number+2, then divided into each reaction tube, and finally RNA samples are added. Reagent 10 µl Reaction system 10×gDNA Remover Mix 1µl RNA Template¹ 10 pg-1 µg RNase-Free Water up to 10 µl Note: 1) If the total RNA amount is greater than 1 µ g, please expand the reaction system proportionally.2. Vortex shake and mix well, briefly centrifuge to collect the solution on the pipe wall to the bottom of the pipe. 3. Incubate at 42 ℃ for 2 minutes (during room temperature reaction, it can be extended to 30 minutes).4. After the reaction is complete, centrifuge briefly and cool on ice.2、 Reverse transcription reaction1. Prepare the reaction system on ice according to the following table, and prepare the reaction solution on ice. To ensure the accuracy of the reaction solution configuration, first prepare a premixed solution in the amount of reaction number+2, and then divide it into 10 batches µ Take 10% of the prepared premix from each reaction tube µ Add to the reaction tube of step 1 that has completed genome removal. Reagent 20 µl Reaction system Step 1 Reaction solution 10µl 5×HiFiScript RTMaster Mix 40µl RNase-Free Water 60µl 2. Mix well and centrifuge briefly to collect the solution on the tube wall to the bottom of the tube.3. cDNA synthesis reaction conditions: Incubate at 37 ℃ for 15 minutes and 85 ℃ for 5 seconds.4. After the reaction is completed, centrifuge briefly and place it on ice before proceeding with the subsequent reaction. If it needs to be stored for a long time, please place it at -20 ℃... Read More | Human PTHrP-(1-36) is a secretory form of PTHrP with anticalciuric effects. Human PTHrP-(1-36) enhances beta cell function and proliferation. Human PTHrP-(1-36) can be used in the research of humoral hypercalcemia of malignancy (HHM) and hyperparathyroidism.In VitroHuman PTHrP-(1-36) (EC 50 : 0.05 Human PTHrP-(1-36) is a secretory form of PTHrP with anticalciuric effects. Human PTHrP-(1-36) enhances beta cell function and proliferation. Human PTHrP-(1-36) can be used in the research of humoral hypercalcemia of malignancy (HHM) and hyperparathyroidism.In VitroHuman PTHrP-(1-36) (EC 50 : 0.05 nM) increases intracellular calcium in human epidermal keratinocytes. Human PTHrP-(1-36) (100 nM, 24 h) increases human β-cell proliferation. Human PTHrP-(1-36) (100 nM, 30 min) enhances insulin secretion in human islets. PTHrP-(1-36) (mouse, EC 50 : 1 nM) induces a rapid Ca 2+ response in UMR 106 cells. MCE has not independently confirmed the accuracy of these methods. They are for reference only.In VivoPTHrP-(1-36) (mouse, 160 µg/kg, s.c., for 5 days/week for 7, 30, or 90 days) enhances beta cell regeneration and increases beta cell mass in a mouse model of partial pancreatectomy. PTHrP-(1-36) (mouse, 100 µg/kg, s.c., every other day) reverses the observed decrease of Wisp1 expression in the diabetic mice. MCE has not independently confirmed the accuracy of these methods. They are for reference only.Form:Solid... Read More | Biochemical Test:SDS-PAGE (purity > 80%); Western blot with patient sample.Calculated Isoelectric Point:pH 5.68 | Malic Dehydrogenase is a ubiquitous enzyme, which exists in two isoforms in eukaryotic cells.Malic dehydrogenase exists as a dimer with each subunit containing an NAD-binding domain and a substrate-binding carboxy-terminal domain required for activity. Malic dehydrogenase is a cytoplasmic isozyme Malic Dehydrogenase is a ubiquitous enzyme, which exists in two isoforms in eukaryotic cells.Malic dehydrogenase exists as a dimer with each subunit containing an NAD-binding domain and a substrate-binding carboxy-terminal domain required for activity. Malic dehydrogenase is a cytoplasmic isozyme and an important catalyst in the tricarboxylic acid cycle.ReagentsA. 0.1 M Tris-HCl buffer (pH7.8)B. 0.01 M Phosphate buffer (KH2PO4-NaOH, pH 7.0)C. Triton X-100 solution (50 mg/ml)D. 0.01 M Phosphate buffer containing 0.1% Triton X-100 (KH2PO4-NaOH, pH 7.0)Dilute 20 ml of Triton X-100 solution (C) with approx. 800 ml of 0.01M Phosphate buffer (B). Fill up to 1,000 ml with 0.01M Phosphate buffer (B).E. NADH soluton Weigh 9 mg of NADH and dissolve in 0.1M Tris-HCl bufer (A). Fill up to 50 ml with 0.1M Tris-HCl Buffer (A). (Can be used for 5 days if kept refrigerated)F. Substrate solutionWeigh 11 mg of oxaloacetic acid and dissolve in 0.1M Tris-HCl buffer (A). Fill up to 50 ml with 0.1M Tris-HCl buffer (A) (Make a fresh solution for each use.)G. Enzyme solutionWeigh out Malate Dehydrogenase and dissolve in chilled 0.01M Phosphate Bufer containing 0.1% Triton X-100 (D). Enzyme solution should be prepared so that the value of AOD/minute becomes in the range of 0.025 ± 0.010.ProcedurePipette 2.0 ml of NADH solution (E) and 0.90 ml of Substrate solution (F) respectively into a quartz cell (d=10 mm) and keep at 25 + 0.5'℃ for 5 minutes. Then, pipete 0.10 ml of Enzyme solution (G) into the quartz cell and mix well immediately. Keep the reaction mixture at 25 ±0.5'C.Exaclly at 2 minutes and 5 minutes after the addition of Enzyme solution (G), measure the absorbances of the reaction mixture at 340 nm(A2 and A5).As a blank, pipette 0.01M Phosphate buffer (D) into another quartz cel (d=10 mm) instead of the Enzyme solution (G) and follow the same procedure described above (Ab2 and Ab5).CalculationMalate dehydrogenase activity (u/mg)=[(A2-A5)-(Ab2-Ab5)]/3*(1/6.22)*(n/0.1) ApplicationThis enzyme is used for the enzymatic determination of L-malate and gluamate oxalo-acetate transaminase(GOT)in clinical diagnosis... Read More | TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP.TMB-D Blotting liquid ready-to-use substrate is a highly active and stable blotting substrate utilized for TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP.TMB-D Blotting liquid ready-to-use substrate is a highly active and stable blotting substrate utilized for measuring HRP probe activity. A stable blue precipitate is formed at the reaction site.The substrate does not contain NMP (1-methyl2-pyrrolidone) making it REACH Restricted Substances List Annex XVII compliant, while ensuring maximal safety during use, and minimal negative environmental impact.Product Characteristics TMB (3, 3', 5, 5'-tetramethylbenzidine) is a chromogenic substrate for Horseradish Peroxidase (HRP). TMB produces a deep blue color during the enzymatic degradation of hydrogen peroxide by HRP.TMB-D Blotting liquid ready-to-use substrate is a highly active and stable blotting substrate utilized for measuring HRP probe activity. A stable blue precipitate is formed at the reaction site. The substrate does not contain NMP (1-methyl-2- pyrrolidone) making it REACH Restricted Substances List Annex XVII compliant, while ensuring maximal safety during use, and minimal waste problems after use.Composition & Properties Ready-to-use substrate: Includes substrate buffer and hydrogen peroxide. No other reagents should be added.Working Procedure The following procedure is applicable to nitrocellulose membranes. The procedure must be optimized for other membranes.1.The desired amount of substrate is poured into a sealed container and allowed to reach room temperature, in the dark, before use. 2.After the last incubation with HRP-labelled Streptavidin or HRP-labelled secondary antibody it is recommended to wash the membrane in a 0.1 M Tris buffer pH 7.4.3.Shake off the excess buffer and incubate the membrane in the TMB-D Blotting solution for 10 minutes. 4.Wash the membrane in distilled water and allow it to dry. 5.The site of positive reaction will appear light blue with no or very little background staining.Tips & Tricks • The membrane can be blocked with Kementec’s Synthetic Blocking Buffer for Blotting, (cat. no. S494457). • For long-term preservation of the results, the membranes must be stored in the dark.Handling & Storage • Store solution at 2-8⁰C in the dark. • Avoid exposure to light, heat and contamination with metal ions or peroxidase. • Re-dispense only into bottles made of High-Density Polyethylene (HDPE), amber color. Dispensing guidelines are available upon request... Read More |