Immunoblotting was performed with rabbit anti-WWP1 antibodies (1 : 700, ab43791; Abcam), and anti-GAPDH antibodies (1 : 1000, ab8245; Abcam) were used as an internal loading control. Membranes were washed and incubated with anti-rabbit IgG horseradish peroxidase-linked antibodies (7074; Cell Signaling Technology, Danvers, MA, USA). Complexes were visualised with Clarity Western ECL Substrate (Bio-Rad, Hercules, CA, USA), as described in our previous studies (Goto et al, 2014a (link); Goto et al, 2015a (link); Kurozumi et al, 2016 (link)).
Ab43791
Manufactured by Abcam
Sourced in United Kingdom
Ab43791 is a primary antibody product offered by Abcam. It is a polyclonal antibody that detects the target antigen. The core function of this product is to bind to and detect the specific target protein in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti rabbit igg horseradish peroxidase linked antibody, by Cell Signaling Technology (1 mentions)
Ab8245, by Abcam (1 mentions)
Clarity western ecl substrate, by Bio-Rad (1 mentions)
Ultravision detection system, by Thermo Fisher Scientific (1 mentions)
Histofine sab po kit, by Nichirei Biosciences (1 mentions)
Electrogenerated chemiluminescence, by Cytiva (1 mentions)
Ab23981, by Abcam (1 mentions)
Ab75285, by Abcam (1 mentions)
Ab181602, by Abcam (1 mentions)
Ab93876, by Abcam (1 mentions)
Anti rabbit antibodies, by Abcam (1 mentions)
Mammalian protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor mixture, by Roche (1 mentions)
3 protocols using ab43791
1
WWP1 Protein Expression Analysis
2
Prostate Tissue Microarray Immunostaining
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A tissue microarray containing a total of 77 prostate specimens was used (Supplementary Table 2 ). The tissue microarray was obtained from Provitro (Berlin, Germany; Cat #401 2209, Lot #146.1 P020212, 26–46). Detailed information on all cancer specimens can be found at http://www.provitro.com/fileadmin/provitro-data/TMA/4012209.pdf . Immunostaining was evaluated using a previously described scoring method (Kojima et al, 2012 (link)). Tissue microarray was immunostained with an Ultra-Vision Detection System (Thermo Scientific, Fremont, CA, USA) following the manufacturer's protocol. Primary rabbit polyclonal antibodies against WWP1 (1:200, ab43791; Abcam) were used for immunochemistry. The slides were treated with biotinylated goat antibodies (Histofine SAB-PO kit; Nichirei, Tokyo, Japan).
3
Western Blot Analysis of Bone Marrow Stromal Cell Proteins
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Whole-cell lysates were prepared from the BMSCs. Cells were lysed using mammalian protein extraction reagent (Pierce Chemical, Dallas, TX), containing protease inhibitor mixture (Roche Applied Science, Indianapolis, IN). The whole-cell lysates (10 μg protein/lane) were loaded and separated in 10% sodium dodecyl sulfate-polyacrylamide gel electropheresis (SDS-PAGE) gels, electro-blotted into a nitrocellulose membrane, and immune-blotted with anti-rabbit antibodies (Abcam, Cambridge, UK) to GAPDH (ab181602, 1:10,000), KLF2 (ab139699, 1:1000), runt-related transcription factor 2 (Runx2, ab23981, 1:1000), osteocalcin (OCN, ab93876, 1:500), osteopontin (OPN, ab75285, 1:1000), WWP1 (ab43791, 1:1000), CD63 (ab134045, 1:1000), CD81 (ab109201, 1:1000), Calnexin (ab92573, 1:20,000), TSG101 (ab125011, 1:1000), p65 (ab32536, 1:1000), phosphorylated p65 (ab86299, 1:2000), IκBα (ab32518, 1:1000), and phosphorylated IκBα (ab133462, 1:10,000). The electrogenerated chemiluminescence (Amersham Biosciences, Piscataway, NJ, USA) was employed to visualize these bands.
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