Rab5a 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.

Rab5 is a 24 kDa GTP-binding protein that regulates the fusion of plasma membrane -derived clathrin-coated vesicles with early endosomes , and homotypic fusion among early endosomes1. Localized to the cytoplasmic side of the plasma membrane, clathrin -coated vesicles and early endosomes, Rab5 appears to regulate vesicle fusion through a cycle of GDP/GTP e xchange and GTP hydrolysis. The different guanine nuc leotide binding states of Rab5 may affect its ability to associate or dissociate with membranes during endocytotic membrane traffic. The GTP-bound, or active, form of Rab5 associates with membrane s and regulates vesicle docking and fusion. Studies using a Rab5 mutant that hydrolyzed xanthosine 5 -triphosphate (XTP) indicated that nucleotide hydrolysis occurs even in the absence of membrane fusion. GTP hydrolysis by Rab5 may determine the frequency of membrane docking and fusion events.

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 NBL1-15082

• Rab5a Antibodies
• Rab5a Lysate
• Rab5a Proteins