Western blot was performed to identify exosomes and verify the predicted proteins associated with B-ALL progression. Whole exosomes in each sample were lysed in radioimmunoprecipitation assay lysis buffer and quantified using a bicinchoninic acid protein assay kit (23,227; Thermo Fisher Scientific). Thereafter, 20 μg of protein was separated by 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred onto a polyvinylidene fluoride membrane (Millipore, Burlington, MA, USA). The membranes were blocked with 5% skim milk and incubated with primary antibodies supplied by Abcam (Cambridge, UK) against TSG101 (ab125011), CD81 (ab109201), calnexin (ab133615), ATG3 (ab108251), and ADAM17 (ab28233) overnight at 4 °C. The membranes were then incubated for 1 h with a secondary antibody at room temperature. A commercial chemiluminescence kit was used to visualize the immunoreactive blots, which were photographed using a Tanon 5200 camera (Tanon, Shanghai, China).
Ab28233
Manufactured by Abcam
Sourced in United Kingdom
Ab28233 is a recombinant monoclonal antibody produced in a mammalian cell line. The antibody is purified and provided in a buffer solution.
Automatically generated - may contain errors
Lab products found in correlation
Ab108251, by Abcam (1 mentions)
Ab125011, by Abcam (1 mentions)
Ab133615, by Abcam (1 mentions)
Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Ab109201, by Abcam (1 mentions)
Ab1827, by Abcam (1 mentions)
Ab191406, by Abcam (1 mentions)
2 protocols using ab28233
1
Exosome Characterization in B-ALL
2
Osteoclast Differentiation and Function
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Unless otherwise stated, all the chemicals and reagents used in this study were supplied by Thermo Fisher Scientific (USA). Antibodies for TIMP-1 (Ab1827), TACE (Ab28233), TRAP (Ab191406), CatK (11239 to 1-AP), and NFATc1 (66963 to 1-Ig) were acquired from Abcam (UK), Proteintech (USA), or Novus Biologicals (USA). Osteoclast Identification kit (70-CK20203), TRAP activity test kit (P0332), F-actin staining kit (C2203-S, Actin-Tracker Red-555), RNeasy Mini Kit (74004), GoScript Reverse Transcription System kit (A5001), and GoTaq qPCR Master Mix kit (A6020) were sourced from MultiSciences (China), Beyotime (China), Qiagen (Germany), and Promega (USA). Enzyme-linked immunosorbent assay (ELISA) kit for TNF-α (10602) was the product of Sino Biological (China). Batimastat (BB-94; S7155), ilomastat (galardin, GM6001; S7157), and TNF-α processing inhibitor (TAPI-0; CAS143457-40-3) were the products of Selleckchem (USA) and Santa Cruz Biotechnology (USA), respectively. Bovine cortical dentin slices (0.2 mm) were purchased from Boneslices (Denmark).
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