| Description | This kit innovatively adopts a column-based purification method to rapidly, gently, and efficiently lyse animal tissues or cells for total protein extraction. It provides both denaturing and native lysis buffers, allowing users to select the appropriate option based on downstream application This kit innovatively adopts a column-based purification method to rapidly, gently, and efficiently lyse animal tissues or cells for total protein extraction. It provides both denaturing and native lysis buffers, allowing users to select the appropriate option based on downstream application requirements. The entire extraction process takes only 1–8 minutes. Thanks to the column purification technology, it can process sample-lysis buffer mixtures as small as 20 µL and up to 500 µL, yielding protein solutions with concentrations of 2–8 mg/mL while effectively preventing protein loss. The extracted proteins can be quantified using the BCA method (Cat. No.: R1491648/B665595). C1491689 Component50TStorageC1491689ADenaturing Lysis Buffer25 mL2-8℃C1491689BNative Lysis Buffer25 mL2-8℃C1491689CProtease Inhibitor Cocktail500 µL-20℃C1491689DPurification Columns50 unitsRT.C1491689ECollection Tubes50 unitsRT.C1491689FPlastic Grinding Pestles4 unitsRT.Key Features1.Simple and rapid operation: Denatured total proteins can be obtained in as little as 1 minute.2.No protein loss: Efficiently extracts DNA-binding proteins by disrupting DNA duplexes.3.Small sample volume, high yield: Processes mixtures as small as 20 µL, yielding protein concentrations of 2–8 mg/mL.4.Versatile applications: Includes two lysis buffers for extracting both denatured and native proteins.Protocol1、Extraction of Denatured Total Protein1.Pre-chill the purification column and collection tube on ice.2.Sample processing: Add protease inhibitor cocktail to the denaturing lysis buffer at a 1:100 ratio shortly before use.2.1 Adherent cells:Wash cells with pre-chilled 1× PBS and aspirate the supernatant.Add the volume of denaturing lysis buffer specified in the appendix table to cover the culture surface, and pipette to mix.2.2 Suspension cells:Collect cells by low-speed centrifugation.Wash with pre-chilled 1× PBS, vortex, and centrifuge at 3,000 rpm for 2–3 minutes.Resuspend the cell pellet in PBS equal to the pellet volume.Add the specified volume of denaturing lysis buffer and vortex to lyse.Note: Partial incomplete lysis does not affect protein extraction. If the lysate is too viscous, directly transfer it to the purification column.2.3 Tissue samples:Place 15–20 mg of tissue on the purification column.Grind 50–60 times with a plastic pestle.Add 200 µL denaturing lysis buffer and grind another 30–60 times.Adjust lysis buffer volume proportionally for larger or smaller samples.Note: Reusable plastic pestles should be thoroughly rinsed with distilled water and dried.3.Centrifugation:3.1 Adherent or suspension cells: Transfer the lysate to the pre-chilled purification column and centrifuge at 14,000–16,000 rpm for 30 seconds.3.2 Tissue samples: Incubate the column at room temperature for 1–2 minutes, then centrifuge at 14,000–16,000 rpm for 1–2 minutes.4.Immediately place the collection tube on ice and discard the purification column. Denatured total protein extraction is complete.2、Extraction of Native Total Protein1.Pre-chill the native lysis buffer, purification column, and collection tube on ice.2.Sample processing: Add protease inhibitor cocktail to the native lysis buffer at a 1:100 ratio shortly before use.2.1 Adherent cells:Wash cells with pre-chilled 1× PBS and aspirate the supernatant.Add the specified volume of native lysis buffer and incubate on ice for 3–5 minutes. Pipette to mix.2.2 Suspension cells:Collect, wash, and resuspend cells as described in section I.Add native lysis buffer, vortex for 15 seconds, incubate on ice for 3–5 minutes, and vortex again for 10 seconds.2.3 Tissue samples:Grind tissue as described in section I, using native lysis buffer.3.Centrifugation:3.1 Adherent or suspension cells: Centrifuge at 14,000–16,000 rpm for 30 seconds.3.2 Tissue samples: Incubate on ice for 5 minutes (open lid), then close the lid and centrifuge at 4°C and 14,000–16,000 rpm for 1–2 minutes.4.Immediately place the collection tube on ice and discard the purification column. Native total protein extraction is complete. Cell Count (×10⁶)Lysis Buffer Volume (µL)0.3200.550110022003500 Appendix: Cell Number vs. Lysis Buffer VolumePrecautionsHigh viscosity of the lysate is normal when using this kit.For safety, wear a lab coat and disposable gloves during operation.For research use only... Read More | DescriptionCAR10 is a kit that contains a selection of 10 carbohydrates/sugars: Arabinose, Fructose, Galactose, Glucose, α-Lactose, Maltose, Mannose, Ribose, Sucrose and Xylose, which may be used for general research, as reagents or as reference compounds in analytical procedures | Product content:E665636Component50 TStorageE665636ABuffer P115 mLRTE665636BBuffer P215 mLRTE665636CBuffer E315 mLRTE665636DBuffer PS15 mLRTE665636EBuffer PW (concentrate)10 mLRTE665636FEndo-Free Buffer EB10 mLRTE665636GRNase A (10 mg/mL)150 µLRTE665636HEndo-Remover FMwith Collection Tubes50 Product content:E665636Component50 TStorageE665636ABuffer P115 mLRTE665636BBuffer P215 mLRTE665636CBuffer E315 mLRTE665636DBuffer PS15 mLRTE665636EBuffer PW (concentrate)10 mLRTE665636FEndo-Free Buffer EB10 mLRTE665636GRNase A (10 mg/mL)150 µLRTE665636HEndo-Remover FMwith Collection Tubes50 EARTE665636ISpin Columns DMwith Collection Tubes50 EART Product Introduction:Endotoxins are a common pollutant in plasmid extraction. Due to the high sensitivity of eukaryotic cells to endotoxins, the presence of endotoxins in plasmids can greatly reduce the transfection efficiency of eukaryotic cells. This reagent kit provides a simple, fast, and efficient new method for extracting endotoxin free plasmids. The extracted plasmids remove endotoxins to the maximum extent possible and can effectively remove contamination of genomic DNA, RNA, proteins, etc. The operation is simple and convenient. This reagent kit is suitable for extracting 1-5mL of bacterial solution. On the basis of alkaline lysis of cells, it efficiently and specifically binds plasmid DNA through a new silicon-based membrane. Each adsorption column can adsorb up to 40% µ The plasmid DNA of g is effectively removed using a special buffer system and endotoxin removal filter column, effectively removing impurities such as endotoxins and proteins. The plasmid obtained from this kit has high purity and stable quality, making it particularly suitable for cell transfection. It can also be used for downstream experiments such as DNA sequencing, PCR, PCR based mutations, in vitro transcription, transformed bacteria, and endonuclease digestion.Self prepared reagents: anhydrous ethanol, isopropanol.Preparation and important precautions before the experiment：1. All components can be stably stored for 1 year in a dry, room temperature (15-30 ℃) environment. The adsorption column can be stored for a longer time at 2-8 ℃. Buffer P1 with RNase A added can be stably stored for 6 months at 2-8 ℃. 2. Before the first use, add all RNase A solution to Buffer P1, mix well, and store at 2-8 ℃. Before use, let it sit at room temperature for a period of time. After returning to room temperature, use.3.Before the first use, anhydrous ethanol should be added to the Buffer PW according to the instructions on the reagent bottle label.4. Before use, please check if there is any crystallization or precipitation in Buffer P2 and Buffer E3. If there is any crystallization or precipitation, you can take a water bath at 37 ℃ for a few minutes to restore clarity.5. Be careful not to come into direct contact with Buffer P2 and Buffer E3, and immediately cover them tightly after use.6.The amount and purity of plasmid extraction are related to factors such as bacterial culture concentration, strain type, plasmid size, and plasmid copy number.Operation steps：1. Take 1-5 mL of overnight cultured bacterial solution and add it to a centrifuge tube (provided). Centrifuge at 13000 rpm (~16200 × g) for 30 seconds to collect bacteria, and try to discard all the supernatant as much as possible.2. Add 250 to the centrifuge tube containing bacterial sediment µ L Buffer P1 (please check if RNase A has been added first), mix thoroughly with a pipette or vortex oscillator, and suspend bacterial precipitation.Attention: If the bacterial blocks are not thoroughly mixed, it will affect the cracking effect, resulting in low extraction amount and purity.3. Add 250 to the centrifuge tube µ L Buffer P2, gently invert and mix 8-10 times, allowing the bacterial cells to fully lyse. Leave at room temperature for 3-5 minutes. At this point, the solution should become clear and viscous.Attention: Mix gently and do not shake vigorously to avoid interrupting genomic DNA and mixing genomic DNA fragments in the extracted plasmid. If the solution does not become clear, it indicates that the bacterial count may be too high and the lysis may not be complete. The bacterial count should be reduced.4. Add 250 to the centrifuge tube µ L Buffer E3, immediately invert and mix 8-10 times until white flocculent precipitates appear. Let it stand at room temperature for 5 minutes. Centrifuge at 13000 rpm for 5 minutes, extract the supernatant, and add it to a filter column (Endo Remove FM). Centrifuge at 13000 rpm for 1 minute to filter, and collect the filtrate in a centrifuge tube (self provided).Attention: After adding Buffer E3, it should be mixed evenly immediately to avoid local precipitation. 5. Add 225 to the filtrate µ Mix L isopropanol upside down.6. Column balance: Add 200 to the spin columns DM that have been loaded into the collection tube µ L Buffer PS, centrifuge at 13000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.7. Transfer the mixed solution of filtrate and isopropanol from step 5 to an equilibrium adsorption column (already loaded into a collection tube).8.13000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.Attention: The maximum volume of the adsorption column is 750 µ L. If the sample volume is greater than 750 µ L can be added in batches. 9. Add 750 to the adsorption column µ L Buffer PW (please check if anhydrous ethanol has been added first), centrifuge at 13000 rpm for 1 minute, and discard the waste liquid in the collection tube.10. Place the adsorption column back into the recovery manifold and centrifuge at 13000 rpm for 1 minute. Note: The purpose of this step is to remove residual ethanol from the adsorption column, which can affect subsequent enzymatic reactions (such as enzyme digestion, PCR, etc.).11. Place the adsorption column in a new collection tube and add 50-100 to the middle of the adsorption membrane µ L Endo Free Buffer EB, let it stand at room temperature for 2-5 minutes, centrifuge at 13000 rpm for 2 minutes, and collect the plasmid solution into a centrifuge tube- Store the plasmid at 20 ℃.Note: 1) To increase the efficiency of plasmid recovery, the obtained solution can be added back to the adsorption column, left at room temperature for 2-5 minutes, centrifuged at 13000 rpm for 2 minutes, and collected into a centrifuge tube.2) When the plasmid copy number is low or>10 kb, preheating the Endo Free Buffer EB in a water bath at 65-70 ℃ can increase the extraction efficiency... Read More | functional group：carboxylic acid Description:Liposome Kit has been used for the preparation of liposomes. Composition：Cholesterol, 9 µmol/package L-α-Phosphatidylcholine (egg yolk), 63 µmol/package Stearylamine, 18 µmol/package | The content of this cell is too long for an XLSX file (more than 32767 characters). Please use the CSV format for this export |