Immunohistochemistry on capillaries were performed as described previously [3 (link), 8 (link), 12 (link)]. The fixed capillaries were incubated with 0.7 µg/mL anti-P-glycoprotein antibody (C219, ThermoFisher Scientific, Rochester, NY) or 1.2 µg/mL anti-BCRP antibody (BXP53, Enzo Life Sciences, Farmingdale, NY) or 1.0 µg/mL anti-MRP2 antibody (M2III-6, Santa Cruz Biotechnology, Dallas, TX) or 10 µg/mL anti-NFkB antibody (ab16502, Abcam, Cambridge, MA) in PBS overnight at 4°C. After rinsing with PBS, capillaries were incubated with Alexa Fluor®568 goat anti-mouse IgG secondary antibody for P-glycoprotein and MRP2 or Alexa Fluor®568 goat anti-rat for BCRP or Alexa Fluor®488 goat anti-rabbit for NFkB (Life Technologies, Grand Island, NY) diluted 1:1000 in PBS, for 90 min at 37°C. Nuclear staining was performed in some of the samples using DRAQ5 (Cell Signaling Technology, Beverly, MA). Immunostaining images of transporters and light contrast images of capillary morphology were captured using Zeiss LSM 510 confocal microscopy.
M2iii 6
Manufactured by Santa Cruz Biotechnology
The M2III-6 is a laboratory instrument designed for the analysis and characterization of biological samples. It is capable of performing various applications, including spectroscopy and fluorescence detection. The core function of the M2III-6 is to provide accurate and reliable data for research and diagnostic purposes.
Automatically generated - may contain errors
Lab products found in correlation
Draq5, by Cell Signaling Technology (1 mentions)
Lsm 510 confocal microscopy, by Zeiss (1 mentions)
Anti gapdh antibody, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase hrp conjugated goat anti mouse antibody, by Santa Cruz Biotechnology (1 mentions)
Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
2 protocols using m2iii 6
1
Immunohistochemical Analysis of Transporter Proteins
2
Western Blot Analysis of MRP2 Expression
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The cells were lysed in RIPA buffer with proteinase inhibitor. After centrifugation, the supernatant was used for western blot analysis. Proteins were separated by SDS-PAGE (8%) and transferred to nitrocellulose membranes; the membranes were incubated with anti-MRP2 monoclonal antibodies M2III-6 (1:200; sc-59,608; Santa Cruz Biotechnology, Dallas, TX; a mouse monoclonal antibody raised against a C-terminal region of MRP2 of human origin) or anti-GAPDH antibodies (1:5000; Santa Cruz Biotechnology) overnight at 4 °C, followed by incubation with horseradish peroxidase (HRP)-conjugated goat anti-mouse antibody (1:5000 dilution; Santa Cruz Biotechnology) for 1 h at 37 °C. Immunocomplexes on the membrane were visualized using Immobilon Western Chemiluminescent HRP Substrate (Millipore, Billerica, USA) and Image Lab Software (BIO-RAD, Hercules, USA).
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