Mitochondria fractionation kit

Manufactured by Beyotime

Sourced in China

The Mitochondria Fractionation Kit is a laboratory equipment designed to isolate and purify mitochondria from various cell types or tissues. The kit provides a standardized protocol and necessary reagents to efficiently separate mitochondria from other cellular components, allowing for further analysis and characterization of this organelle.

Automatically generated - may contain errors

Lab products found in correlation

Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions) 5 ethynyl 2 deoxyuridine edu incorporation assay kit, by RiboBio (1 mentions) Pde2 activity assay kit, by Abcam (1 mentions) Bay 60 7550, by MedChemExpress (1 mentions) Pka kinase activity assay kit, by Abcam (1 mentions) P8340, by Merck Group (1 mentions) Rapi lysis buffer, by Beyotime (1 mentions) Caspase 3 p17, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Mitochondria/Cytosol Fractionation Kit (27 citations)

7 protocols using mitochondria fractionation kit

Mitochondrial Fractionation and Enzyme Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

The primary antibodies and their working concentrations used in this study are listed in Supplementary Material Table 1.2.3. The Mitochondria Fractionation Kit (Cat# C3601) was purchased from Beyotime (Biotechnology, China). PDE2 Activity Assay Kit (Cat# ab139460), PKA Kinase Activity Assay Kit (Cat# ab139435), cAMP Assay (Cat# ab65355) and ODQ (Cat# ab120022) were all purchased from Abcam (Abcam, United Kingdom). Go 6983 (Cas# 133053-19-7) inhibitor was from MCE (MCE, USA). The CellTiter 96^® Aqueous One Solution Cell Proliferation Assay was purchased from Promega (Cat# G3581, Promega, USA) and 5-ethynyl-2’-deoxyuridine (EdU)-incorporation assay kit was purchased from Ribobio (Cat# C10310, Ribobio, China). Bay 60-7550 (Cat# HY-14992) and H89 (Cat# HY-15979) were purchased from Med Chemexpress (Med Chemexpress, USA).

Zhao Y., Wang Y., Zhao J., Zhang Z., Jin M., Zhou F., Jin C., Zhang J., Xing J., Wang N., He X, & Ren T. (2021). PDE2 Inhibits PKA-Mediated Phosphorylation of TFAM to Promote Mitochondrial Ca2+-Induced Colorectal Cancer Growth. Frontiers in Oncology, 11, 663778.

+ Open protocol

+ Expand

Isolation of Hippocampal Mitochondria

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mitochondria of hippocampus were isolated using Mitochondria Fractionation Kit (Beyotime) as previously described (Du et al., 2016 (link)). Briefly, the hippocampus tissues were quickly removed from adult male offspring and placed in chilled isolation media (0.25 M sucrose, 10 mM Tris–HCl buffer, pH 7.4, 1 m MEDTA, and 250 μg BSA/ml). The tissues were minced and washed with the isolation medium, and 10% (w/v) homogenates were ready. Nuclei and cell debris were sedimented by centrifugation at 600 g for 10 min at 4°C and discarded. The supernatant was subjected to centrifugation at 10,000 g for 10 min at 4°C. The resulting mitochondrial pellets were suspended in the isolation medium.

Wu T., Huang Y., Gong Y., Xu Y., Lu J., Sheng H, & Ni X. (2019). Treadmill Exercise Ameliorates Depression-Like Behavior in the Rats With Prenatal Dexamethasone Exposure: The Role of Hippocampal Mitochondria. Frontiers in Neuroscience, 13, 264.

+ Open protocol

+ Expand

Mitochondria Isolation from Heart Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

The mitochondria were isolated using a Mitochondria Fractionation Kit (Beyotime Biotech, Shanghai, P.R. China) based on a previously reported method.³² (link) The extracted heart tissues were swiftly placed in mannitol/sucrose/HEPES/EGTA (MSHE) buffer containing 0.22 M mannitol, 70 mM sucrose, 0.5 mM ethylene glycol-bis (β-aminoethyl ether)-N, N-tetraacetic acid (EGTA), and 2 mM K-N-(2-hydroxyethyl) piperazine-N’-(2-ethanesulfonic) acid (HEPES), at 4°C. The myocardium samples were subsequently minced into homogenates, which were then centrifuged at 700 rpm for 10 min to remove the debris. Next, the collected supernatant was centrifuged at 1,000 rpm for 10 min. Finally, the mitochondrial pellets were collected for follow-up experimentation.

Zuo T., Tang Q., Zhang X, & Shang F. (2020). MicroRNA-410-3p Binds to TLR2 and Alleviates Myocardial Mitochondrial Dysfunction and Chemokine Production in LPS-Induced Sepsis. Molecular Therapy. Nucleic Acids, 22, 273-284.

+ Open protocol

+ Expand

Mitochondrial Fractionation and Subfractionation

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mitochondria and cytosol fractions from mouse liver or glutamine-treated HeLa cells or hippocampal slices were isolated using a mitochondria fractionation kit from Beyotime Biotechnology (China) according to manufacturer’s instructions. Mitochondrial proteins and cytosolic proteins were used for Western blotting. Mitochondrial subfractionation assay was performed as previously described (Li et al., 2016 (link); She et al., 2011 (link)). Isolated mitochondria were resuspended in 10 µM KH₂PO₄ (pH 7.4) for 20 min on ice. An equal volume of iso-osmotic solution (32% sucrose, 30% glycerol, 10 mM MgCl₂) was added and spun at 10,000 g and 4°C for 10 min. The supernatant was centrifuged at 15,000 g and 4°C for 1 h; the pellet and supernatant contained outer membrane (OM) and intermembrane space (IMS) proteins. Then, the pellet was resuspended in 10 µM KH₂PO₄ (pH 7.4) for 20 min on ice, and iso-osmotic solution was added, followed by centrifugation at 15,000 g and 4°C for 1 h; the pellet and supernatant contained inner membrane (IM) and matrix (Mx) proteins.

Yang W., Pang D., Chen M., Du C., Jia L., Wang L., He Y., Jiang W., Luo L., Yu Z., Mao M., Yuan Q., Tang P., Xia X., Cui Y., Jing B., Platero A., Liu Y., Wei Y., Worley P.F, & Xiao B. (2021). Rheb mediates neuronal-activity-induced mitochondrial energetics through mTORC1-independent PDH activation. Developmental cell, 56(6), 811-825.e6.

+ Open protocol

+ Expand

Isolation and Assessment of Mitochondria

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isolation of mitochondria was performed using the Mitochondria Fractionation Kit (Beyotime, China). Briefly, mouse lung tissues were quickly removed and placed in beakers containing chilled (4°C) isolation media (0.25M sucrose, 10mM Tris–HCl buffer, pH 7.4, 1mM EDTA and 250μg BSA/ml. The tissues were minced and washed three times with the isolation media to remove adhering blood and 10% (w/v) homogenates were prepared using homogenizer. The nuclei and cell debris were sedimented by centrifugation at 600g for 10min and discarded. The supernatant was subjected to a further centrifugation at 10000g for 10min. Mitochondrial pellets were suspended in the isolation medium. Respiratory control ratio (RCR) was used to assess the quality of isolated mitochondria.

Huang Y., Wang G., Zhou Z., Tang Z., Zhang N., Zhu X, & Ni X. (2021). Endogenous Hydrogen Sulfide Is an Important Factor in Maintaining Arterial Oxygen Saturation. Frontiers in Pharmacology, 12, 677110.

+ Open protocol

+ Expand

Cardiac Mitochondria Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isolation of mitochondria was performed with the Mitochondria Fractionation Kit (Beyotime Biotech, Haimen, China) according to the manufacturer's protocol. Briefly, fresh cardiac tissue was mixed with a mitochondria extraction reagent and stirred in a homogenizer and the suspension was centrifuged at 1000 × g for 5 min. (4°C); the supernatant obtained was centrifuged at 3500 × g for 10 min. (4°C) and the precipitate contained the mitochondrial fraction.

Xue R., Sun L., Yu X., Li D, & Zang W. (2016). Vagal nerve stimulation improves mitochondrial dynamics via an M3 receptor/CaMKKβ/AMPK pathway in isoproterenol‐induced myocardial ischaemia. Journal of Cellular and Molecular Medicine, 21(1), 58-71.

+ Open protocol

+ Expand

Protein Extraction and Analysis in Liver

Check if the same lab product or an alternative is used in the 5 most similar protocols

For total protein extraction, liver tissues or cell samples were lysed in RAPI lysis buffer (P0013, Beyotime) containing protease inhibitors (P8340, Sigma) according to the manufacturer’s instructions. Nuclear components were extracted according to Cytoplasmic and Nuclear Protein Extraction Kit (BB-36021, Bestbio). Isolation of mitochondrial proteins was performed using the Mitochondria Fractionation Kit (C3603, Beyotime Inst. Biotech). This study used the antibodies against the following proteins: SHP (PA5-102494, Invitrogene); GAPDH (60004-I-Ig, Proteintech); Sox9 (SC-166505), Bax (sc-7480), Cyt C (sc-13156), Bid (sc-373939) and Caspase-3 p17 (sc-271028) from Santa Cruz.

Qin D., Wang R., Ji J., Wang D., Lu Y., Cao S., Chen Y., Wang L., Chen X, & Zhang L. (2023). Hepatocyte-specific Sox9 knockout ameliorates acute liver injury by suppressing SHP signaling and improving mitochondrial function. Cell & Bioscience, 13, 159.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!