U-2 OS Nuclear Cell Lysate

• Reactivity: Hu
• Applications: WB, IP
• Concentration: 2 mg/ml

U-2 OS Nuclear Cell Lysate Summary

• Description: Quality control test: 12.5% SDS-PAGE Stained with Coomassie Blue. Nuclear extract cell lysate (non-denatured)
• Localization: Bone
• Specificity: U-2 OS (human osterogenic sarcoma) 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.

Applications/Dilutions

• Dilutions:
  • Immunoprecipitation
  • Western Blot
• 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.

Packaging, Storage & Formulations

• 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

Lysate Details for Array

• Type: Cell
• Tissue: Bone/Cartilage
• Protein State: Native
• Tissue Condition: Osteogenic Sarcoma
• Subcellular Fraction: Nuclear

Notes

This product is produced by and distributed for Abnova, a company based in Taiwan.

Background

U2OS is a human bone osteosarcoma from a human 15 year old Caucasian female expressing wild type p53 and lacking p16. U2OS also has a ZFN modification creating a GFP-NUP98 transgene expressed from the endogenous NUP98 gene locus. U2OS cells were established in 1964, the original cells were taken from a moderately differentiated sarcoma of the tibia. U2OS cells are positive for insulin-like growth factor I (IGF-I) and insulin-like growth factor II (IGF II) receptors and express a number of antigens, including blood type A, Rh+, HLA A2, Aw30, B12, Bw35, and B40(+/-).

Limitations

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