FBXW7/Cdc4 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Auxin-inducible degron (AID) technology allows rapid depletion of proteins in animal cells and fungi. Auxin includes chemicals such as IAA (indole-3-aceticacid) and NAA (1-naphtalene acetic acid) and includes inhibitors in plants. For the degradation, and SCF ubiquitin ligase complex containing the TIR1 protein is activated via the association of auxin with TIR1. Because the core SCF components are conserved in all eukaryotic cells, it is possible to transplant the degradation pathway to yeast and mammalian cells by expressing TIR1. In those cells expressing TIR1, a protein fused with degron (AID degron) derived from AUX/IAA can be rapidly degraded in the presence of auxin. Anti-mini-AID tag can detected both mini-AID-tag and full-length AID-tag.

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.

Array NBP2-11238

• FBXW7/Cdc4 Antibodies
• FBXW7/Cdc4 Lysates
• FBXW7/Cdc4 Proteins