Raw 264.7 Nuclear Cell Lysate

Product Discontinued

Raw 264.7 Nuclear Cell Lysate Summary

• Description: Quality control test: 12.5% SDS-PAGE Stained with Coomassie Blue. Nuclear extract cell lysate (non-denatured)
• Localization: macrophage
• Specificity: Raw 264.7 (mouse macrophage, Abelson murine leukemia virus transformed) 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: Blood Lymph
• Protein State: Native
• Tissue Condition: Abelson Leukemia virus transformed
• Subcellular Fraction: Nuclear

Notes

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

Background

Raw264.7 cells are a macrophage-like, Abelson leukemia virus transformed cell line derived from BALB/c mice. They were established from the ascites of a tumor induced male mouse via intraperitoneal injection of Anselon Leukemia Virus (A-MuLV). Raw264.7 cells are capable of antibody dependent lysis of sheep erythrocytes and tumor targets. Growth of Raw264.7 cells is inhibited by lipopolysaccharides (LPS).

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.