Skip to main content
Fig. 5 | Microbial Cell Factories

Fig. 5

From: Design and validation of recombinant protein standards for quantitative Western blot analysis of cannabinoid CB1 receptor density in cell membranes: an alternative to radioligand binding methods

Fig. 5

Quantitative Western blot analysis of CB1 receptor density on P2 membranes from rat brain cortex. A, D, G Increasing amounts of total protein from P2 membranes were loaded together with increasing molar amounts of combined GST/GST-CB1414-442 or GST/GST-CB1414-472 recombinant proteins, resolved side by side on SDS-PAGE and processed for immunoblotting using CB1-Af380 (A), CB1-Af450 (D) and CB1-ImmGs (G) polyclonal antibodies, all raised against the 31 amino acid sequence at the C-terminus of the CB1 receptor. B, E, H Immunoreactive bands profile detected in P2 samples of brain cortex from adult CB1-WT and CB1-KO mice were used to identify the CB1 receptor-specific signals detected by CB1-Af380 (C), CB1-Af450 (F) and CB1-ImmGs (I) antibodies. C, F, I Slopes obtained by linear regression analysis of the integrated optical density (Integrated OD) values of immunoreactivity corresponding to the endogenous CB1 receptor in P2 membranes and to the recombinant construct GST-CB1414-472. The integrated OD values were obtained by densitometric analysis of the immunoreactive bands produced by the endogenous CB1 receptor at ~ 50 and ~ 35 kDa and by the recombinant standard GST-CB1414-472 (subtracting the value of the non-specific signal produced by the GST-CB1414-442 protein) at increasing sample loads

Back to article page

Microbial Cell Factories

ISSN: 1475-2859

Contact us