Equal amounts of EVs or total cell lysates were separated on 4–12% bis‐Tris gels using MES buffer (Thermo) and transferred to PVDF membranes (Immobilon‐P, Millipore). The membrane was blocked in 5% milk in TBS with 0.1% Tween 20, incubated with primary antibodies (mouse anti‐Alix, Cell Signalling 2171; mouse anti‐Tsg101, Genetex GTX70255; rabbit anti‐ovalbumin, Novus NB600‐922‐01; rabbit anti‐Rab27a, Cell Signalling 69295; mouse anti‐b‐catenin, Millipore Mab2081), washed, and incubated with corresponding secondary antibodies (sheep anti‐mouse‐HRP, GE Healthcare NA931 V; goat anti‐rabbit‐HRP, Dako P0448) before signal development using ECL substrate and ECL Hyperfilm (both GE Healthcare).
Mab2081
Manufactured by Merck Group
Mab2081 is a laboratory equipment product offered by Merck Group. It is designed for use in various research applications, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach without further interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Hyperfilm ecl, by GE Healthcare (1 mentions)
Goat anti rabbit hrp, by Agilent Technologies (1 mentions)
Mes buffer, by Thermo Fisher Scientific (1 mentions)
Immobilon p, by Merck Group (1 mentions)
Mouse anti alix, by Cell Signaling Technology (1 mentions)
Mouse anti tsg101, by GeneTex (1 mentions)
Prolong antifade reagent, by Thermo Fisher Scientific (1 mentions)
Acti stain 555 fluorescent phalloidin, by Cytoskeleton (1 mentions)
2 protocols using mab2081
1
Western Blot Analysis of EVs
2
Immunofluorescence Staining of CXCR6 and β-Catenin
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Subconfluent cells grown on glass coverslips are fixed with 3.7% parafolmaldeide in PBS for 10min, permealized with 0.5% Triton X-100 in PBS for 5min at room temperature and incubated with 10% goat serum in PBS for 1hr. The cells are stained with polyclonal anti-CXCR6 antibody (1:400, Abcam, Ab8023) or anti-β-catenin (MIllipore, MAB2081, 1:200 nostro; anti-Active βCatenin (Anti-ABC) Antibody, Millipore, 05-665, 1:50) or Anti-Fascin antibody (DyLight 488, ab156578 Abcam) overnight at 4 °C. Thus, after a brief washing with PBS, CXCR6 staining cells are incubated with the secondary antibody (anti rabbit Alexa488 1:250) for 1 h. The cytoskeleton is stained with Falloidin (Acti-stain 555 Fluorescent Phalloidin, Cytoskeleton Inc, 100 nM) at room temperature for 45 min. The nuclei are counterstained with DAPI and the slides mounted with Pro-long anti fade reagent (Life technologies). The images are acquired with a Leika TCS NT confocal microscope.
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