Minute™ Plasma Membrane Protein Isolation and Cell Fractionation Kit (50 Preps)
Manual & Protocol (PDF) | Material Safety Data Sheets (MSDS)(PDF)
View All Related Products
For orders and inquiries
The MinuteTM kit represents the next generation product for plasma membrane (PM) isolation and cell fractionation, offering unsurpassed convenience and consistency by eliminating uncontrollable variations associated with traditional homogenization and density gradient centrifugation including two phase partitioning.
How it works: cells/tissues are first sensitized by buffer A before passing through the proprietary filter in a zigzag manner when high-speed centrifugal force is applied, resulting in a cell lysate containing ruptured cell membranes and intact nuclei. As a result, the nuclear contaminations are virtually eliminated. PM is further separated from the cell lysate (a mixture of crude membranes, intact nuclei, cytosol proteins and organelles) by subsequent differential and density centrifugation with a regular tabletop microcentrifuge. 5 distinct cell fractions (total membrane, PM, cytosol, nucleus and organelles) can be obtained at the completion of the protocol. The procedure can be completed in less than 45 minutes.
Figure 1. A. SDS-PAGE profiles of total cell lysate (TC) and isolated plasma membrane proteins (PM) from human and rat cultured cells. Lane 1, Human lung cancer cell A549 total cell lysate; Lane 2, Isolated PM of A549; Lane 3, PM of rat REL-6TN cells; Lane 4, Total cell lysate of rat REL-6TN cells.
B. Western blotting: Proteins in A were transferred to nitrocellulose membrane and probed with anti-Na/K ATPase alpha1, a plasma membrane marker (Upstate, Clone 464.6) and anti-lamin B1, a nuclear envelope marker (ab16048, abcam Cambridge, MA). The specific protein bands were visualized by a color metric substrate Opti-4CN (Bio-RAD).
C. Densitometry measurement of Na/K ATPase alpha1 signals in B (TC vs. PM). Total cell lysates were extracted by Minute Denaturing Total Protein Extraction Kit (SD-001, Invent Biotechnologies, Eden Prairie, MN).
Figure 2 A. SDS-PAGE profiles of isolated total membrane proteins from mouse tissues (T=Total Cell Lysate, C=Cytosol Fraction, M= Total Membrane Fraction)
B. Proteins shown in A were transferred to a nitrocellulose membrane and probed with rabbit-anti mouse pan-cadherin (ab6529, abcam, Cambridge, MA), and anti-actin by Western blotting. The specific protein bands were visualized by a color metric substrate Opti-4CN (Bio-RAD). Signals of pan-cadherin (a 130 kDa plasma membrane marker) were significantly enhanced in total membrane protein fractions. Total cell lysates were extracted by Minute™ Denaturing Total Protein Extraction Kit (SD-001, Invent Biotechnologies, Eden Prairie, MN).
Figure 3. A. SDS-PAGE profiles of total membrane protein fraction (TM) and isolated plasma membrane protein fraction from mouse tissues.
B. Western blotting of proteins in A were transferred to nitrocellulose membrane and probed with rabbit anti-cadherin (abcam, Cambridge, MA). The specific protein bands were visualized by a color metric substrate Opti-4CN (Bio-RAD).
C. Densitometry measurement of cadherin signals in B (TM vs. PM).
|Items||Quantity||Buffer A||25 ml|
|Buffer B||10 ml|
|Protein Extraction Filter Cartridges||50 units|
|Collection Tubes with Caps||50 units|
|Plastic Rods||2 units|
|Tissue Dissociation Beads||10 grams|
1. Kalabus, JL. et al. (2012). MicroRNAs differentially regulate carbonyl reductase 1 (CBR1) gene expression dependent on the allele status of common polymorphic variant rs9024. PLOS ONE. 7(11):1-9.
2. Lombrafia AQ. et al (2014). Interindividual variability in the cardiac expression of anthracycline reductases in donors with and without down syndrome. Pharm Res. 31:1644-1655.
3. Hou, N., et al. (2014). Vitamin A deficiency impairs spatial learning and memory: The mechanism of abnormal CBP-dependent histone acetylation regulated by retinoic acid receptor alpha. Mol Neurobiol. DOI 10.1007/s12035-014-8741-6.
4. You, Y., et al. (2014). Latexin sensitizes leukemogenic cells to gamma-irradiation-induced cell-cycle arrest and cell death through Rps3. Cell Death and Disease. (2014)5, e1493; doi: 10.1038/cddis. 2014.443.
5. Tanaka M et al. (2016). Ex vivo effects of naphthoquinones allergen-sensitized mononuclear cells in mice. International Journal of Immunopathology and Pharmacology. DOI: 10.1177/0394632016632221
6. Liu W. et al. (2016) Electroacupunctre improves motor impairment via inhibition of microglia-mediated neuroinflammation in the sensorimotor cortex after ischemic stroke. Life Sciences. doi:10.1016/j.lfs.2016.01.045
7. Xu, Yang, et al. (2016), RIP3 induces ischemic neuronal DNA degradation and programmed necrosis in rat via AIF." Scientific Reports 6:29362.
8. Shi, Le, et al. (2016). "Evodiamine exerts anti-tumor effects against hepatocellular carcinoma through inhibiting β-catenin-mediated angiogenesis." Tumor Biology: 1-13.9
9. Matsuoka, Y., Nakayama, H., Yoshida, R., Hirosue, A., Nagata, M., Tanaka, T., ... & Shinriki, S. (2016). IL-6 controls resistance to radiation by suppressing oxidative stress via the Nrf2-antioxidant pathway in oral squamous cell carcinoma. British Journal of Cancer.
10. Ueda, K., Masuda, A., Fukuda, M., Tanaka, S., Hosokawa, M., & Iwakawa, S. (2017). Monophosphorylation by deoxycytidine kinase affects apparent cellular uptake of decitabine in HCT116 colon cancer cells. Drug Metabolism and Pharmacokinetics.
11. Kawaguchi, R., Hirata, N., Tokinaga, Y., Hayashi, S., Inoue, S., Watanabe, N., & Yamakage, M. (2017). Nitrite administration improves sepsis-induced myocardial and mitochondrial dysfunction by modulating stress signal responses. Journal of Anesthesia, 31(6), 885-894.
12. Kong, P., Yu, Y., Wang, L., Dou, Y. Q., Zhang, X. H., Cui, Y., ... & Cui, W. (2019). circ-Sirt1 controls NF-κB activation via sequence-specific interaction and enhancement of SIRT1 expression by binding to miR-132/212 in vascular smooth muscle cells. Nucleic Acids Research.
13. Kang, Y., Li, Y., Zhang, T., Chi, Y., & Liu, M. Effects of transcription factor EB on oxidative stress and apoptosis induced by high glucose in podocytes. International Journal of Molecular Medicine. 44(2), 447-456.
14. Yuan, H. X., Chen, C. Y., Li, Y. Q., Ning, D. S., Li, Y., Chen, Y. T., ... & Jian, Y. P. (2020). Circulating extracellular vesicles from patients with valvular heart disease induce neutrophil chemotaxis via FOXO3a and the inhibiting role of dexmedetomidine. American Journal of Physiology-Endocrinology adoi.org/10.1007/s13402-020-00540-6nd Metabolism.
15. Dang, H., Chen, L., Tang, P., Cai, X., Zhang, W., Zhang, R., ... & Tang, H. (2020). LINC01419 promotes cell proliferation and metastasis in hepatocellular carcinoma by enhancing NDRG1 promoter activity. Cellular Oncology, 1-17.
We're here to help
Get expert recommendations for common problems or connect directly with an on staff expert for technical assistance related to applications, equipment and general product use.
Contact Tech Support
High Quality Guaranteed Product
Our products such as Elisa, Antibodies, Proteins, Peptides and sequencing kits are covered by Biolinkk quality warranty and will work as described in datasheet, a free replacement or money back is guaranteed if does not perform according to datasheet.