mitochondrial extraction kit
Cat.No：SM0020 Solarbio
Storage：Short-term store at 2-8℃，1 months;Long-term store at -20℃.
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PI3K/Akt/mTOR
Endocrinology & Hormones
Protein Tyrosine Kinase/PTK
Angiogenesis
HIF
Immunology & Inflammation
Stem Cells
Apoptosis
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ER Stress
Cell Cycle
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DNA Damage & DNA Repai
Epigenetics
Ubiquitin
JAK/STAT
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Antibody-drug Conjugate/ADC Related
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Pipette & Dispenser & Pipette Dispensing Instrument
Oscillating Mixing & Stirring & Dispersing Instrument
Temperature Control & Sampling & Grinding Crushing Instrument
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PCR Instrument & Light Glue Instrument
Spectrophotometer
Centrifuge
Shaker
Vacuum Pump
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Product Information
| Name | mitochondrial extraction kit |
| Storage | Short-term store at 2-8℃，1 months;Long-term store at -20℃. |
| Transport Condition | The product is shipped on ice pack. |
| Unit | Kit |
| Specification | 50T 100T |
Product Introduction:
Mitochondrial extraction kits are used to isolate complete and purified mitochondria from animal cells or tissues. Suitable for the preparation of animal soft tissues (liver or brain tissue) and hard tissues (muscle) as well as mitochondria of cultured cells. It has a high yield and can be used for the study of apoptosis, signaling, metabolism and proteomics.
![/></p><p><b> Operation steps: </b></p>A. Tissue homogenate: Weigh 100~200 mg of fresh tissue such as liver, brain, myocardium, etc., rinse with PBS or normal saline, wash off blood water, blot dry with filter paper, cut into pieces with scissors and put them into a small-capacity glass homogenizer. Lysis Buffer with 1.0 mL ice was added and the lysis was grinded up and down for 20 times in a 0℃ ice bath. b. Cultured cell homogenate: digested cells, washed with PBS, centrifuged at 800g for 5-10 min, collected cells and counted them. The Lysis Buffer of 5×10<sup>7</sup> cells was added to 1.0 mL of suspended cells pre-cooled by ice for each extraction. The cell suspension was transferred to a small-capacity glass homogenizer and ground in a 0℃ ice bath for 30~40 times. </p><p>2. Transfer the tissue or cell homogenate to a centrifuge tube and centrifuge at 1000g at 4℃ for 5 min. </p><p>3. Remove the supernatant and transfer it to a new centrifuge tube. Centrifuge again at 1000g at 4℃ for 5 min. </p><p>4. Remove the supernatant and transfer it to a new centrifuge tube, centrifuge at 4℃ at 12,000 g for 10 min. The supernatant after centrifugation contains cytoplasmic components from which cytoplasmic proteins can be extracted. The supernatant is transferred to a new centrifuge tube, and the mitochondria are deposited on the bottom of the tube. </p><p>5. 0.5mL Wash Buffer was added to the mitochondrial precipitation to suspend the mitochondrial precipitation, and centrifuged at 1000 g at 4℃ for 5 min. </p><p>6. Take the supernatant and transfer it to a new centrifuge tube, centrifuge at 4℃ at 12,000 g for 10 min. After the supernatant is discarded, high purity mitochondria are deposited on the bottom of the tube. </p><p>7. Re-suspend mitochondrial precipitation with 50-100 μL Store Buffer or appropriate reaction buffer for immediate use or -70℃ preservation. </p><p><b> Note: </b></p><p>1. In order to obtain complete mitochondria, it is necessary to: first, the whole process of low temperature operation; Second, fast; Third, breaking up cells without destroying suborganelles is the most critical step in making mitochondria. Compared with tissue blocks, cultured cells, especially adherent cells, are more difficult to break the wall when homogenized by glass homogenizer, so small capacity glass homogenizer and tight gap pestle should be used to grind the cultured cells up and down. </p><p>2. Calculate the correct centrifugal speed with centrifugal force g, and different centrifuges can accurately calculate the centrifugal speed accordingly. 3. Western Blot and 2D-gel electrophoresis were performed, and the sample buffer could be directly added to split mitochondria. </p><p> Rotational speed and centrifugal force conversion: </p><p>G = 1.11×(10-5)×R×[rpm]<sup>2</sup>G is the centrifugal force, generally expressed as a multiple of g (acceleration of gravity); [rpm]<sup>2</sup> : the square of the speed; R is the radius in centimeters. </p><p><b>Note：<span style=](/images/upload/image/20160615/20160615160117_27539.png)
Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.
Remark：These protocols are for reference only. Solarbio does not independently validate these methods.
Note：
1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.
2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.
3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.
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