| Reactivity | HuSpecies Glossary |
| Applications | WB, IP |
| Concentration | 2 mg/ml |
| Description | Quality control test: 12.5% SDS-PAGE Stained with Coomassie Blue. Nuclear extract cell lysate (non-denatured) |
| Localization | Epidermis |
| Specificity | A-431 (human epidermoid carcinoma) nuclear extract lysate (non-denatured) |
| Preparation Method |
Nuclear extract was prepared by using a modified protocol of Dignam et al. Cells were Harvested and homogenized in Buffer A, and then centrifugated at 25,000 g for 20 minutes to remove cytoplasm and pellet the nuclei. The pellet was re-suspended in Buffer C, and then the suspensions were centrifuged to collect nuclear extract. The supernatant was dialyzed against Buffer D. The dialysate was then centrifuged, divided into aliquots, and stored at -80C. The protein concentration was determined by the method of Bradford. The lysate was adjusted to 2 mg/ml. |
| Dilutions |
|
| Application Notes | Use it directly for immuno-precipitation, or heat lysate with SDS gel loading buffer to 95C for 5 minutes followed by rapid cooling for western blot application. If dissociating conditions are required, add reducing agent prior to heating. |
| Storage | Store at -80C. Avoid freeze-thaw cycles. |
| Buffer | 20mM HEPES pH 7.9, 20%(v/v) glycerol, 50mM KCl, 0.2 mM EDTA, 0.5 mM DTT & 0.5 mM PMSF. |
| Concentration | 2 mg/ml |
| Type | Cell |
| Tissue | Skin |
| Protein State | Native |
| Tissue Condition | Epidermoid Carcinoma |
| Subcellular Fraction | Nuclear |
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
