Nuclear extraction kit instructions
Nuclear extraction kit instruction manual article number: SN0020
Specification: 50T/100T
Storage: Store at 4°C for short-term use and -20°C or -70°C for long-term storage. product content:
Component SN0020 -50 SN0020-100
Lysis Buffer 100 mL 200 mL
Reagent A 2.5mL 5mL
Medium Buffer 25 mL 50 mL
Store Buffer 5 mL 10 mL
Product introduction:
A nuclear extraction kit is used to isolate intact and purified nuclei from animal cells or tissues. Suitable for animal soft tissues (liver or brain tissue) and hard tissue (muscle) as well as the preparation of the nuclei of cells in culture. Its preparation yield is high and can be used for research on apoptosis, signaling, metabolism and proteomics. The product does not contain contaminating proteases and ribozymes and has stable performance.
Cell nuclear extraction kit steps: 1. Sample processing
a tissue homogenate: Weigh 100 ~ 200 mg fresh tissue such as the liver, brain, heart, etc., PBS or normal saline, wash bloody, dry filter paper, cut with scissors into small-capacity broken piece of glass homogenizer Inside. Add 1.0 mL of pre-cooled Lysis Buffer, add 50ul of LReagent A, and grind the tissue up and down 20 times in an ice bath at 0 °C ; there are unground tissue, which can be filtered with double gauze. (b) culturing the cell homogenate: cells were digested, washed with PBS, 800 g centrifuged at 5 ~ 10 min the cells were collected, counted. Each extraction required 5 × 107 cells, were added 1.0 mL of ice-cold Lysis Buffer resuspended cells, then add 50ul Reagent A, the cell suspension was transferred to a small glass homogenizer capacity, 0 ℃ ice bath was triturated cell 20~30 times. 2. Transfer tissue or cell homogenate to a 1.5 ml centrifuge tube and centrifuge at 700 g for 5 min at 4 °C . The nucleus was deposited at the bottom of the collection tube and the supernatant was discarded. Resuspend the pellet by adding 0.5 mL of ice-cold Lysis Buffer. 3. Add 0.5 mL of Medium Buffer to another new centrifuge tube, pipette the resuspension from the previous step, carefully add it to the tube along the tube wall, place it on the Medium Buffer, and centrifuge at 700 g for 5 min at 4 °C . The nucleus is deposited at the bottom of the tube. 4. Discard the supernatant, resuspend the nuclear pellet by adding 0.5 mL Lysis Buffer in the nuclear pellet, centrifuge at 1000g for 10min, discard the supernatant, and obtain a pure nuclear pellet. 5. Resuspend the nuclear pellet with 50-100 μL of Store Buffer or a suitable reaction buffer and store immediately or at -70 °C.
Nuclear extraction kit notes:
1. To ensure complete nuclei sure that: first, the whole low temperature operation; a second, fast; third, broken cells without damaging the subcellular organelles of the case, which is the most critical part of the preparation of the nucleus. Compared with the tissue block, the cultured cells, especially the adherent cultured cells, are more difficult to break when homogenized with a glass homogenizer . Therefore, the cells should be cultured up and down by using a small-capacity glass homogenizer and a well-stitched mortar.
2. The centrifugal force g calculate the correct speed centrifugation, the centrifuge may be different centrifugation speed accurately calculated accordingly.
3. Perform Western Blot and 2D-gel electrophoresis, and directly add the loading buffer to lyse the nucleus. Speed of the centrifugal force transducer calculated: G = 1.11 × (10-5) × R × [rpm] 2 G of centrifugal force, typically by a factor G (gravitational acceleration) is represented; [rpm] 2 that is: the square of the rotational speed; is R & lt Radius in centimeters.
Related Products:
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SM0020 Mitochondrial Extraction Kit
Preservation tubes are swabs with disposable virus sampling tubes to collect DNA tests for disposable nasal flocking sterile medical transport.
Made of high-quality PP material, high chemical stability, high temperature sterilization, suitable for multi-channel pipettes and automatic machines. Square well, U-shaped bottom design ensures minimal sample residue. Molded letters and numbers for easy positioning. Complies with SBS regulations and can be stacked stably. Good sealing performance, the top of the plate is flat and uniform, maintaining an effective seal. Optional gamma radiation sterile, DNase and RNA enzyme free, pyrogen-free.
Stable performance, low price, the material has a strong electrostatic adsorption capacity, can provide customized plastic card color according to user requirements, good color consistency
Non-invasive, painless and easy to operate. Samples are non-contagious and can be transported safely.
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Jiangsu iiLO Biotechnology Co., Ltd. , https://www.iilogene.com