Twenty micrograms of total cellular and EV lysates were separated by SDS-PAGE before transferring to PVDF membrane (Pall Australia, VIC, Australia), as described in [4 (link)]. The membrane was blocked overnight with 5% skim milk in PBS and 0.05% Tween 20 and then incubated with anti-P-gp mAb (clone F4; Sigma-Aldrich), anti-Ezrin mAb (clone 3C12; Life Science), anti-Moesin mAb (clone 38/87; Sigma-Aldrich), anti-Radixin pAb (SAB2500859; Sigma-Aldrich) or anti-CD44 pAb (HPA005785; Sigma-Aldrich) for 1 h. Anti-β-actin (clone AC-74; Sigma-Aldrich) was used as the internal control, followed by 1 h incubation with anti-Mouse HRP secondary antibody (Promega, NSW, Australia) at a 1:10,000 dilution. Protein expression was visualised using the ECL (enhanced chemiluminescence) system (Roche Applied Science, Castle Hill, NSW, Australia). The membranes were imaged using the luminescent image analyser LAS-3000 (Fujifilms, Brookvale, NSW, Australia).
Anti p gp mab clone f4
Manufactured by Merck Group
Sourced in Australia
The Anti-P-gp mAb (clone F4) is a monoclonal antibody that specifically binds to the P-glycoprotein (P-gp) efflux transporter. It can be used as a tool in research and development applications to study the expression and functional activity of P-gp.
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Lab products found in correlation
2 protocols using anti p gp mab clone f4
1
Western Blot Analysis of Membrane Proteins
2
Quantification of Membrane Protein Expression
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Twenty micrograms of total cellular and MP proteins were separated on SDS-PAGE, as described earlier, before transferring to PVDF membrane (Pall Australia, VIC, Australia). The membrane was blocked overnight with 5% skim milk in PBS and 0.05% Tween 20, and then incubated with anti-P-gp mAb (clone F4; Sigma-Aldrich), anti-Ezrin mAb (clone 3C12; Life Science), anti-Moesin mAb (clone 38/87; Sigma-Aldrich), anti-radixin pAb (SAB2500859; Sigma-Aldrich) or CD44 mAb (clone EPR1013Y; Abcam) for 1 h. Anti-β-actin (clone AC-74; Sigma-Aldrich) was used as the internal control, followed by horseradish peroxidase conjugates secondary antibody. Protein expression was visualized using an ECL (enhanced chemoluminescence) system (Roche Applied Science, NSW, Australia). The membranes were imaged using the luminescent image analyser LAS-3000 (Fujifilms, Brookvale, NSW, Australia).
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