Nuclear Extract Kit
for high quality nuclear, cytoplasmic or whole-cell extract preparation
The Nuclear Extract Kit isolates high-quality nuclear, cytoplasmic or whole-cell extract from mammalian cell or tissue samples. This eliminates the need for optimizing reagents and protocols, and ensures high yields every time. More importantly, standardizing your extraction procedure with our high-quality reagents ensures more reproducible results. Extracts are suitable for use in a various applications including EMSA, Western blot and binding assays and is ideally suited for preparation of extracts for our TransAM® Transcription Factor Assays.
NEK Fractionation Western blots
The Nuclear Extract Kit works with fresh or frozen cell and tissue samples and yields protein amounts of approximately 0.5 to 1 mg from cytoplasmic extract (at 1-2 mg/ml concentration), 150 to 250 μg from nuclear extract (at 3-5 mg/ml concentration), 1.2 to 2.4 mg from whole cell extract (at 4-8 mg/ml concentration) from 8.8 x 106 cells. The kit procedure is simple and only takes two hours to complete. The kit includes buffers, detergents and reagents for lysis as well as Protease and Phosphatase inhibitors to ensure preservation of your proteins.
To learn more about our Nuclear Extract Kit, click on the Description or Contents tabs below. To view a manual or other related documents, click on the Documents tab below.
|Nuclear Extract Kit||100 rxns||40010||$235||Buy Now|
|400 rxns||40410||$690||Buy Now|
|Nuclear Extract Manual|
|Nuclear Extract Profile|
|Cell Biology Products Brochure|
|Gene Regulation Products & Services Brochure|
Extracts prepared using the Nuclear Extract Kit can be used with Active Motif’s TransAM® Kits, or in other procedures that require cell extracts, like gelshift assays, Western blots, DNA footprinting, as a starting point for protein purification for binding or activity assays or or other analysis techniques.
Nuclear Extract Kit advantages
- Quality-controlled reagents ensure reproducibility
- No need to optimize your own procedure
- Complete kit contains all required reagents
- Prepare nuclear, cytoplasmic or whole-cell extracts with one kit
To demonstrate the efficiency of cellular fractionation using the Nuclear Extract Kit, nuclear, cytoplasmic, and whole-cell extracts were prepared from HeLa cell samples. Nuclear pellets from the nuclear extraction were also collected. Cellular fractions were analyzed by Western blot using antibodies against various proteins. As shown in Figure 1, proteins that serve as markers for cellular fractionation, such as nuclear marker RNA pol II and cytoplasmic marker alpha Tubulin, were only detected in their respective cellular fractions and in whole-cell extracts. The localization of other proteins, such as membrane-associated Lamin A/C and pan-Cadherin, mitochondrial COX IV and GAPDH were also analyzed.
Figure 1: Western blots of specific proteins from cellular fractions obtained with the Nuclear Extract Kit.
To demonstrate the quality of the materials and protocol of the Nuclear Extract Kit, nuclear and cytoplasmic extracts were prepared from HeLa cell samples that were either unstimulated or stimulated with TNF-α for 30 minutes. Because activated NFκB translocates into the nucleus, only nuclear extract from stimulated cells should contain activated NFκB. As predicted, this is precisely what was seen (Figure 2), demonstrating the kit’s high specificity.
Figure 2: Specific extraction of nuclear and cytoplasmic extracts.
Active Motif also offers a number of ready-to-use nuclear, cytoplasmic and whole-cell extracts. These extracts have been prepared using the same high-quality components found in the Nuclear Extract, and many are supplied as positive controls in TransAM Transcription Factor DNA-binding ELISAs.
Contents & Storage
Lysis Buffer, Dithiothreitol, Protease Inhibitor Cocktail, 10X PBS, Phosphatase Inhibitors, 10X Hypotonic Buffer and Detergent. Reagent storage conditions vary from 4°C to -20°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly