DORFIN 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.

Dorfin is an E3 ubiquitin ligase which mediates ubiquitination of cellular proteins. Dorfin contains two RING-finger motifs and an IBR (in between RING fingers) motif. Dorfin interacts with UBE2L3/UBCH7 and UBE2E2/UBCH8, but not other ubiquitin-conjugating enzymes. Dorfin is of interest in Parkinson's Disease, as it binds and ubiquitylates synphilin 1 (SNCAIP - interacts with alpha synuclein) in neurons. Furthermore, Dorfin expression is increased in Lewy bodies, the characteristic neuronal inclusions in Parkinson's diseased brains. In normal subjects, Dorfin is widely expressed at low levels with higher levels found in the heart and is ubiquitously expressed in the central nervous system. Alternatively spliced transcript variants encoding the same protein have been reported.

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 NBP2-04770

• DORFIN Antibodies
• DORFIN Lysates
• DORFIN Protein