Subcellular Protein Fractionation Kit

Subcellular Protein Fractionation Kit [NBP2-47659]

• Reactivity: Ma
• Applications: WB, Enzyme Activity, GS

Subcellular Protein Fractionation Kit Summary

• Description: The Cell Fractionation kit is designed to provide reproducible extraction of four subcellular protein fractions (cytosol, nucleus, membrane/particulate, and cytoskeletal fractions) from a single mammalian sample. The method is fast and simple, needing only 2 hours and no ultracentrifugation involved. All four protein fractions obtained are suitable for many downstream applications such as 1-D or 2-D gel, enzyme activity assays, gel shift assay, and Western blotting.
  
  The percentage of proteins in the different fractions are cell-dependent, usually in the range of 20-30% cytosol, 25-35% membrane (organelles), 15-25% Nuclei, 15-25% insoluble.
  
  This product is manufactured by Abcam and distributed by Novus Biologicals. (Abcam Catalog Number: ab288085)
• Kit Type: Fractionation Kit

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
  • Gel Supershift Assay
  • Western Blot
• Application Notes: In Western analysis, if a membrane protein appeares mainly in the nuclear fraction, the membrane extraction step may not be sufficient. Make sure to add Membrane Extraction Buffer B into the extraction, which contains NP-40 detergent. If it still shows the same result, increase incubation time from 1 min to 5 min after adding MEB-B. NP-40 should extract membrane proteins from most of cells.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Publications for Subcellular Protein Kit (NBP2-47659)(3)

Publications using NBP2-47659

• Luguang Liu, Longgang Wang, Liqing Liu, Xianlin Qu, Weizhu Zhao, Jishuang Ding, Siwei Zhao, Botao Xu, Hang Yu, Bing Liu, Jie Chai Acyltransferase zinc finger DHHC-type containing 2 aggravates gastric carcinoma growth by targeting Nrf2 signaling: A mechanism-based multicombination bionic nano-drug therapy. Redox biology 2024-02-26 [PMID: 38301594]
• Xu M, Tan J, Zhu L et al. Palmitoyltransferase ZDHHC3 Aggravates Nonalcoholic Steatohepatitis by Targeting S-Palmitoylated IRHOM2 Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2023-08-06 [PMID: 37544908]
• Blazejewski SM, Bennison SA, Ha NT Et al. Rpsa Signaling Regulates Cortical Neuronal Morphogenesis via Its Ligand, PEDF, and Plasma Membrane Interaction Partner, Itga6 Cerebral cortex (New York, N.Y. : 1991) 2021-08-04 [PMID: 34347028]

Review for Subcellular Protein Kit (NBP2-47659) (1)

Average Rating: 4

(Based on 1 review)

reviewed by: Babak Nami 10/23/2018

Summary

• Lot: 4E09K02700

Comments

• Comments: This kit works very well mammalian cell lines. The procedure is quite simple and fast. Results in very clean membrane, cytosol and nuclear fractionation but cytoskeletal fraction is contaminated by nuclear fraction. The only problem is that it seems that all fractions are contaminated by cytosol. I got a lot of a-tubulin in the all fractions!!! Very strong a-tubulin band in membrane fraction. a-tubulin is supposed to be cytosolic protein. Having it in cytoskeletal fraction makes sense but not in membrane and nuclear fraction. In my manual protocol I used to use I get tubulin-free membrane and nuclear fractions.