| Description | Inquire | Description：Acetylcholinesterases (AChEs) are enzymes that hydrolyze the neurotransmitter acetylcholine (ACh) to acetate and choline. AChE is located at the synaptic cleft and functions to terminate synaptic transmission by catalyzing the breakdown of ACh allowing cholinergic neurons to Description：Acetylcholinesterases (AChEs) are enzymes that hydrolyze the neurotransmitter acetylcholine (ACh) to acetate and choline. AChE is located at the synaptic cleft and functions to terminate synaptic transmission by catalyzing the breakdown of ACh allowing cholinergic neurons to return to a resting state after activation. It is also found in membranes of red blood cells, motor and sensory fibers, muscles, nerves and central and peripheral tissues. Changes in AChE activity may result from exposure to certain insecticides, which act as cholinesterase inhibitors. Inhibitors of AChE are also used to treat certain conditions such as dementia.Acetylcholinesterase activity assay kit has been used to determine the activity of acetylcholinesterase in a rat organophosphate model and in brain tissue homogenates.Principle：Acetylcholinesterase can catalyze the hydrolysis of acetylcholine to choline, and the reaction of choline with disulfide p-nitrobenzoic acid to produce 5-merhydryl-nitrobenzoic acid (TNB). The product has a characteristic absorption peak at 412 nm, and the activity of acetylcholinesterase can be characterized by the change of light absorption valueThe Dilution Calculator EquationConcentration (start)xVolume (start)= Concentration (final)× Volume (final)This equation is commonly abbreviated as: C1V1 = C2V2... Read More | Products contentN665968Component96 TStorageN665968Adex N501-N508 Primers for Illumina 8×12 µL-20℃. Avoid freeze/thaw cycle.N665968BIndex N701-N712 Primers for Illumina 12×8 µL-20℃. Avoid freeze/thaw cycle. Products IntroductionThis kit is a companion kit for the Products contentN665968Component96 TStorageN665968Adex N501-N508 Primers for Illumina 8×12 µL-20℃. Avoid freeze/thaw cycle.N665968BIndex N701-N712 Primers for Illumina 12×8 µL-20℃. Avoid freeze/thaw cycle. Products IntroductionThis kit is a companion kit for the transposase-based second-generation sequencing Rapid DNA Library Construction Kit, designed for Illumina platform library construction, which contains 8 primers at the N5 end and 12 primers at the N7 end, which can be used to prepare 96 different bipartite Index libraries. All reagents provided in the kit have been subjected to stringent quality control and functional validation to maximize the stability and reproducibility of library construction. The prepared libraries can be sequenced on Illumina platforms such as HiSeq X-10/4000/2500/2000 and MiSeq. Provide your own instruments, reagents and consumables1. Magnetic frame: DynaMagTM-2 is recommended.2. DNA purification and recovery kit: It is recommended to use Kangwei DNA purification and recovery kit by magnetic bead method.3. DNA building kit: It is recommended to use the Kangwei Century transposase method second-generation sequencing rapid DNA building kit.4. Anhydrous ethanol.5. Reaction tubes: It is recommended to use low adsorption PCR tubes and 1.5 ml centrifuge tubes; tips: It is recommended to use high-quality filtration tips to prevent contamination of reagent kits and library samples. Pre-experiment Preparation and Important NotesPlease centrifuge briefly before opening the cap so that the liquid collects at the bottom of the tube to avoid cross-contamination between different primers.For the use of the CombiVision Second Generation Sequencing Multisample Primer Kit, please follow the CombiVision Second Generation Sequencing Rapid DNA Library Kit protocol.Index N501-N508 Primers for IlluminaIndex N701-N712 Primers for Illumina... Read More | Products R669890Component50 TStorageR669890ADNase I1000 U-20℃. Avoid freeze/thaw cycle.R669890B10×Reaction Buffer1mL-20℃. Avoid freeze/thaw cycle.R669890CBuffer RL35 mLRTR669890DBuffer RW140 mLRTR669890EBuffer RW2 (concentrate)11 mLRTR669890FRNase-Free Water10 mLRTR669890GSpin Products R669890Component50 TStorageR669890ADNase I1000 U-20℃. Avoid freeze/thaw cycle.R669890B10×Reaction Buffer1mL-20℃. Avoid freeze/thaw cycle.R669890CBuffer RL35 mLRTR669890DBuffer RW140 mLRTR669890EBuffer RW2 (concentrate)11 mLRTR669890FRNase-Free Water10 mLRTR669890GSpin Columns FL with Collection Tubes50 setsRTR669890HSpin Columns RM with Collection Tubes50 setsRTR669890IRNase-Free Centrifuge Tubes (1.5 mL)100 EART ProductsThis kit adopts centrifugal adsorption columns with high efficiency and specificbinding of nucleic acids and unique buffer system, which can rapidly extract totalRNA from bacteria or cultured animal cells.The reaction can be completed in 30-40minutes, and the extracted total RNA is extremely pure and free of protein and othercontaminants, which is suitable for RT-PCR, Real-Time RT-PCR, microarray analysis,in vitro translation and other experiments. Self-contained reagents: Lysozyme, β-mercaptoethanol, anhydrous ethanol (freshlyopened or for RNA extraction). Pre-experiment Preparation and Important Notes 1. To prevent RNase contamination, attention should be paid to the following aspects:1) Use RNase-free plastics and tips to avoid cross-contamination. 2) RNase-free water should be used to prepare the solution. 3) Operators wear disposable masks and gloves, and change gloves diligently duringthe experiment. 2. Add β-mercaptoethanol to Buffer RL before use to reach a final concentrationof 1%, e.g., add 10 µl of β-mercaptoethanol to 1 ml of Buffer RL. Buffer RL withβ-mercaptoethanol can be stored at 4℃ for 1 month, if precipitation occurs, pleaseheat to dissolve and use.3. Anhydrous ethanol should be added to Buffer RW2 before first use according tothe instructions on the reagent bottle label. 4. All centrifugation steps are carried out at room temperature if not otherwisespecified, and all steps should be performed quickly. Procedure 1. Centrifuge at 12,000 rpm (~13,400 x g) at 4°C for 2 minutes to collect theorganisms (maximum volume of organisms should not exceed 1 x 109) and carefullyremove all supernatants. Note: Supernatants that leave residues can interfere with the subsequent digestionprocess. 2. Thoroughly resuspend the organisms with 100 µl of TE buffer containing Lysozymeand incubate at room temperature. The specific formulation and incubation time areas follows:/The final concentration of Lysozyme in TE bufferincubation timeG-germ400µg/ml3-5minG+germ3mg/ml5-10min 3. Add 350 µl of Buffer RL (check that β-mercaptoethanol has been added beforeuse), vortex and shake to mix (insoluble precipitate may appear in this step), addall of the solution and the precipitate to the filter columns (Spin Columns FL) thathave been loaded into the collection tubes, and centrifuge at 12,000 rpm for 2minutes. 4. Add 250 µl of anhydrous ethanol to the filtrate obtained in the previous stepand mix well (a precipitate may appear at this point). Transfer the resulting solution together with the precipitate to a Spin Columns RM packed in a collectiontube, centrifuge at 12,000 rpm for 1 min, discard the waste solution and put thecolumn back into the collection tube.5. Add 350 µl Buffer RW1 to the adsorbent column, centrifuge at 12,000 rpm for1min, discard the waste liquid and put the adsorbent column back into the collectiontube.6. Preparation of DNase I mixture: Take 52µl of RNase-Free Water, add 8µl of 10×Reaction Buffer and 20µl of DNase I (1U/µl) to it, mix well, and make a finalvolume of 80µl of reaction solution.7. Add 80µl of DNase I mixture directly to the adsorption column and incubate at20-30°C for 15 minutes.8. Add 350 µl Buffer RW1 to the adsorbent column, centrifuge at 12,000 rpm for1min, discard the waste liquid and put the adsorbent column back into the collectiontube.9. Add 500 µl of Buffer RW2 to the column (check that anhydrous ethanol is addedbefore use), centrifuge at 12,000 rpm for 1 min, and discard the waste solution.10. Repeat step 9.11. Place the adsorbent column back into the collection tube and centrifuge at 12,000rpm for 2 minutes. Note: The purpose of this step is to remove residual ethanol from the adsorptioncolumn; ethanol residue can interfere with subsequent enzymatic reactions (zymography, PCR, etc.).12. Load the adsorption column into a new RNase-Free collection tube, add 30-50 µl of RNase-Free Water to the middle of the adsorption membrane, leave it at roomtemperature for 1 minute, centrifuge at 12,000 rpm for 1 minute, collect the RNAsolution, and store the RNA at -70°C to prevent degradation. Note: 1) The volume of RNase-Free Water should not be less than 30 µl, too smallvolume affects the recovery rate. 2) If you want to increase the RNA yield, repeat step 12 with 30-50 µl of freshRNase-Free Water. If the RNA concentration is to be increased, the resulting solution can be reintroduced into the adsorption column and step 12 repeated... Read More | Product content R669871Component50 TStorageR669871ADNase I1000 U-20℃. Avoid freeze/thaw cycle.R669871B10×Reaction Buffer1mL-20℃. Avoid freeze/thaw cycle. R669871CBuffer DS30 mLRTR669871DBuffer GTL15 mLRTR669871EBuffer GL25 mLRTR669871FProteinase K12.5 mgRTR669871GProteinase K Product content R669871Component50 TStorageR669871ADNase I1000 U-20℃. Avoid freeze/thaw cycle.R669871B10×Reaction Buffer1mL-20℃. Avoid freeze/thaw cycle. R669871CBuffer DS30 mLRTR669871DBuffer GTL15 mLRTR669871EBuffer GL25 mLRTR669871FProteinase K12.5 mgRTR669871GProteinase K Storage Buffer1.25 mLRTR669871HBuffer RW140 mLRTR669871IBuffer RW2 (concentrate)11 mLRTR669871JRNase-Free Water10 mLRTR669871KSpin Columns RS with Collection Tubes50 setsRTR669871LRNase-Free Centrifuge Tubes (1.5 mL)50 EART Product IntroductionThis kit is suitable for effectively purifying total RNA from formalin fixed and paraffin embedded tissues. Suitable for extracting total RNA with improved purity from paraffin embedded tissues or sections less than 30mg. This kit does not require the use of phenol/chloroform extraction or isopropanol precipitation, and can complete the extraction of multiple samples within one hour. This product uses specially optimized lysis solution and protease K to release RNA from formalin fixed or tissue slice samples without overnight operation; After digestion, the sample is incubated at a higher temperature to remove the inhibitory effect caused by formalin cross-linking, effectively releasing RNA from tissue slices and avoiding endangering RNA integrity; The optimized buffer system allows RNA in the lysis solution to specifically bind to the silica gel adsorption membrane, while other pollutants can flow through the membrane; It can be effectively removed through rinsing steps, and the washed RNA can be directly used for experiments such as RT-PCR, Real Time PCR, and Western blot analysis.Self prepared reagents: anhydrous ethanol (newly opened or dedicated for RNA extraction), 10mM PBS (pH 7.4).Preparation and important precautions before the experiment1. Add 0.625ml Protein K Storage Buffer to Protein K to dissolve it and store at -20 ℃. The prepared Protein K should not be left at room temperature for a long time to avoid repeated freeze-thaw cycles, which may affect its activity.2. To prevent RNase pollution, attention should be paid to the following aspects:1) Use RNase free plastic products and gun heads to avoid cross contamination.2) Glassware should be dry baked at a high temperature of 180 ℃ for 4 hours before use, while plastic containers can be soaked in 0.5M NaOH for 10 minutes, thoroughly rinsed with water, and then sterilized under high pressure.3) Prepare the solution using water without RNase.4) Operators should wear disposable masks and gloves, and change gloves frequently during the experiment.3. After obtaining the sample, it should be fixed in 4% -10% formalin as soon as possible, with a suitable fixation time of 14-24 hours. Excessive time can lead to RNA breakage and affect downstream experiments.4. Ensure that the sample before embedding is thoroughly dehydrated, as residual formalin will inhibit the action of Protein K.5. Before the first use, anhydrous ethanol should be added to Buffer RW2 according to the instructions on the reagent bottle label.Before use, please check if there is any crystallization or precipitation in Buffer GTL, Buffer GL, and Buffer DS. If there is any crystallization or precipitation, please dissolve Buffer GTL, Buffer GL, and Buffer DS again in a 56 ℃ water bath.Operation steps1. Sample processing1a. Paraffin embedded sample: Use a surgical knife to trim off excess paraffin from the tissue block, expose the tissue, and cut into 5-10 µ m thin slices.Attention: If the surface of the sample has already been exposed to air, please discard 2-3 pieces that come into contact with the air and do not use them.1b. Samples in fixed solutions such as formalin: Take approximately 20mg of the sample, cut it into small pieces, place it in a centrifuge tube, and add 500 µ 10mM PBS (PH7.4), vortex oscillation, centrifugation at 12000 rpm (~13400 × g) for 1 minute, discard the supernatant, repeat 3 times, and proceed directly to step 3.2. Choose option A or option B to remove paraffinOption AA1. Take approximately 1 × 1cm2 of slices (4-5 slices in total) and place them in a centrifuge tube (prepared by oneself), then add 500 slices µ L Buffer DS, vortex oscillation for 10 seconds. Incubate at 56 ° C for 3 minutes.Centrifuge at A2.12000 rpm for 2 minutes, be careful to discard the supernatant and avoid attracting sediment.Option BB1. Take approximately 4-5 slices of approximately 1 × 1 cm2 and place them in a centrifuge tube (self prepared). Add 1ml of xylene, cover the tube tightly, and vortex for 10 seconds.B2.Centrifuge at 12000 rpm for 2 minutes, be careful to remove the supernatant and avoid removing sediment.B3. Add 1ml of anhydrous ethanol, vortex and shake well. Centrifuge at 12000 rpm for 2 minutes, discard the supernatant, and be careful not to absorb or discard the sediment.B4. Open the tube cover and incubate at room temperature or up to 37 ° C for 10 minutes until there is no ethanol residue.3. Add 150µ L Buffer GTL, resuspended precipitation; Join 10µl Protein K, vortex oscillation mixing.4.Incubate at 56 ℃ for 15 minutes until the sample is completely dissolved. Incubate at 80 ℃ for 15 minutes. Short centrifugation allows the solution on the tube wall to be collected to the bottom of the tube.Note: 1) The purpose of this step is to repair nucleic acids denatured by formaldehyde. Incubating at a high temperature or for too long may cause RNA breakage, resulting in RNA fragments.2) The sample incubated at 56 ℃ can be placed at room temperature until the temperature of the water or dry bath reaches 80 ℃, and then the sample can be incubated at 80 ℃.5. Place on ice for 3 minutes, centrifuge at 12000 rpm for 15 minutes, transfer the supernatant to a new centrifuge tube, be careful not to suck sediment.6. Add 320 to the supernatant µ L Buffer GL, vortex oscillation thoroughly mixed.7. Join 720 µ Mix anhydrous ethanol thoroughly with vortex oscillation.Attention: After adding anhydrous ethanol, there may be a small amount of precipitate precipitation, but it does not affect subsequent operations.8. Add all the solutions obtained in step 7 to the spin columns RS that have been loaded into the collection tube. If the solution cannot be added at once, it can be transferred multiple times. Centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.Optional steps: If genomic DNA needs to be removed, the following steps can be followeda. Add 350 to the adsorption column µ L Buffer RW1, centrifuge at 12000 rpm for 1 minute, discard the waste liquid, and place the adsorption column back into the recovery manifold.b. Preparation of DNase I mixture: Take 52 µ Add 8 RNase Free Water to it µ 10 x Reaction Buffer and 20 µ DNase I (1U/ µ l) Mix well and prepare to a final volume of 80 µ The reaction solution of L.c. Add 80 µ l of DNase I mixture directly to the adsorption column and incubate at 20-30 ℃ for 15 minutes.d. Add 350 to the adsorption column µ L Buffer RW1, centrifuge at 12000 rpm for 1 minute, discard the waste liquid, and place the adsorption column back into the recovery manifold.9. Add 500 to the adsorption column µ Buffer RW2 (check if anhydrous ethanol has been added before use), centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.10. Repeat step 9.Centrifuge at 11.12000 rpm for 2 minutes and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes to thoroughly air dry.Note: The purpose of this step is to remove residual ethanol from the adsorption column, which will affect subsequent enzymatic reactions (such as enzyme digestion, PCR, etc.).12. Place the adsorption column in a new RNase free centrifuge tube, and add 20-50µl to the middle of the adsorption column in the air Place RNase Free Water at room temperature for 2-5 minutes, centrifuge at 12000 rpm for 1 minute, collect RNA solution, and store RNA at -20 ℃.Note: 1) The volume of RNase Free Water should not be less than 20 µ l. Small volume affects the recovery rate. 2) If you want to increase RNA production, you can use 20-50 µ Repeat step 12 for the new RNase Free Water.3) If you want to increase the RNA concentration, you can add the obtained solution back to the adsorption column and repeat step 12... Read More |