Nuclear Complex Co-IP Kit

Produktnummer ABIN2866218

Produktdetails

Applikation: Immunoprecipitation (IP)

Spezifität: Nuclear Complex Co-IP Kits are quality control tested to demonstrate that they are effective in performing co-immunoprecipitation experiments with nuclear protein complexes.

Anwendungsinformationen

Kommentare:

HeLa cells are grown to confluence on 100 mm plates and nuclear extracts prepared using the kit's extraction reagents. For IP experiments, both IP High and IP Low Buffers were used in parallel with the p33/RNApol II Co-IP. The stringency of the IP High Buffer was increased by supplementing with NaCl (150 mM Final) and Detergent (1% Final). The stringency of the IP Low Buffer was unchanged. 100 μg of nuclear extract was used per IP reaction and incubated with either 2 μg p33 antibody or no antibody (negative control). Protein G beads were then added to each IP reaction. Following the wash steps of the IP, 2X reducing sample buffer was added to each IP reaction, and samples were boiled and run on an SDS-PAGE gel. Western blot analysis was performed using RNA pol II mouse mAb at 0.1 μg/ml followed by anti-mouse HRP at 1:1000. Detection of the p33/RNA pol II complex by the RNA pol II antibody (lane 3) demonstrates that the Co-IP was successful in maintaining the protein complex. The input HeLa extract (lane 1) was run as a control for the western blot using 0.1 μg/ml RNA pol II. The negative (no antibody) control was run as a control for the IP reaction.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: The Nuclear Complex Co-IP Kit contains many different components that must be stored at either room temperature, 4°C or -20°C. Please consult the manual for the different storage conditions for each individual component. This product is guaranteed for 6 months from date of receipt.

Haltbarkeit: 6 months