Rab4 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.

Rab4 is a small GTPase that regulates recycling of proteins, such as GPCRs, from the early endosomes to the cell surface. Overexpression of Rab4 causes a redistribution of receptors on plasma membrane versus endocytic compartments. Rab4 is also involved in the translocation of glucose transporter (Glu4) in adipocytes in response to insulin. Association of Rab4 with transferrin receptor containing early endosomes is mediated through the geranylgeranyl groups at its carboxyl terminus. Rab4 can be used as a marker for early endosomes (see Daro et al. 1996 and Zuk et al. 1999 for more information). Membrane association is also cell cycle dependent, as phosphorylation at its C-terminal cdc2 kinase consensus sequence in mitotic cells leads to dissociation of Rab4 into the cytosol.

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 NBL1-15080

• Rab4 Antibodies
• Rab4 Lysate
• Rab4 Protein