EVs-specific proteins were analyzed using the capillary Western blot Jess system (ProteinSimple, CA, USA) according to the manufacturer’s protocol. Briefly, samples were diluted to a concentration of 1 mg/mL, denatured at 95 °C for 5 min, and processed using the 12–230 kDa Jess Separation Module with different antibodies: anti-TSG101 antibody (Novus Biologicals LLC, Cat. NB200-112, 1/10), anti-CD63 antibody (R&D Systems; Cat. MAB50482, 1:20), anti-calnexin antibody (Novus Biologicals LLC; Cat. NB100-1965, 1:25). The fluorescence detection was quantified with streptavidin-HRP (ProteinSimple; Cat. 042–414). After protein migration (25 minutes at 375 V) they were incubated for 30 minutes with the primary antibodies, followed by a washing step and a 30-minute incubation with the secondary antibodies. Finally, the Compass Simple Western software (version 5.0.1, ProteinSimple) was used to capture the digital image of the capillary immunodetection.
Anti cd63 antibody
Manufactured by R&D Systems
The Anti-CD63 antibody is a laboratory research tool used to detect and study the CD63 protein, which is a membrane glycoprotein involved in various cellular processes. The antibody can be used in techniques such as flow cytometry, immunohistochemistry, and Western blotting to identify and analyze CD63-expressing cells and tissues.
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Lab products found in correlation
2 protocols using anti cd63 antibody
1
Protein Analysis of Extracellular Vesicles
2
Quantification of Tumor Markers CA125 and CA15-3
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The 96‐well microtiter plates coated with streptavidin (SA plates, product #: 41‐07TY), assay wash buffer (product #: 42‐01TY), and RED assay buffer (product #: 42‐02TY) were purchased from Kaivogen Oy (Turku, Finland). From Fujirebio Diagnostics, we acquired the anti‐CA125 antibody Ov197 and anti‐CA15‐3 antibody Ma552 as well as the conventional assay kits for CA125 and CA15‐3. The anti‐CD63 antibody was purchased from R&D Systems (Minneapolis, Minnesota). The plate washer (Delfia PlateWash 1296–026), plate shaker (Delfia PlateWash 1296‐026), and VictorX4™ fluorimeter were from Wallac Oy (Turku, Finland). The reagents for cell line cultures were Gibco brand of Thermo Fisher Scientific (Waltham, Massachusetts) except for glutamine, which was Ultraglutamine from Lonza (Basel, Switzerland), and phosphate buffered saline which was from GE Healthcare (Chicago, Illinois).
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