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Ab81234

Manufactured by Abcam
Sourced in China

Ab81234 is a lab equipment product. It is a device designed for use in scientific research and laboratory settings. The core function of this product is to facilitate specific laboratory procedures or analyses. No further details on the intended use or application of this product can be provided in an unbiased and factual manner.

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3 protocols using ab81234

1

Western Blotting Optimization in S. Typhimurium

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Reagents for Western blotting were obtained from Bio-Rad (Hercules, CA, USA). Proteins were electrophoretically separated on a sodium dodecyl sulfate (SDS)-polyacrylamide gel, followed by transfer to nitrocellulose membranes. The transformation of pcDNA6.2-GW/EmGFP-mi-INHA into S. Typhimurium was assessed by monitoring the co-cistronic expression of EmGFP using a monoclonal anti-GFP antibody (ab1218, Abcam). The expression of bacterial chaperone protein DnaK was probed for simultaneously using an anti-DnaK antibody (ab69617, Abcam) as the internal loading control. The other proteins in mammalian cancer cells were detected using the following primary anti-bodies: INHA (ab81234) and Bcl-2 (ab59348) from Abcam; Bcl-xL (2764) and actin (4967) from Cell Signaling (Beverly, MA, USA). Secondary antibodies were horseradish peroxidase labeled anti-mouse or anti-rabbit antibodies. Binding was detected by chemiluminescence signals, according to the manufacturer’s instructions (Roche, Basel, Switzerland).
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2

Protein Extraction and Western Blotting

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The GCs were harvested and washed once in phosphate-buffered saline (PBS), then lysed on ice for 30 min with radioimmunoprecipitation assay (RIPA) buffer (CST, 9806), and supplemented with 1% (v/v) protease inhibitor Cocktail (HY-K0010) and 1% (v/v) phosphatase inhibitors (Cocktail I, HY-K0021; Cocktail II, HY-K0022; and Cocktail III, HY-K0023), which were purchased from MCE (Shanghai, China). Western blotting was performed as described previously (Wu et al., 2019 (link); Yang et al., 2020a (link)). Protein concentrations were determined using a BCA protein assay kit (TransGen Biotech, Beijing, China). Equal amounts of proteins (15–50 μg/lane) were separated by SDS-PAGE (12% acrylamide running gel) and transferred to a nitrocellulose membrane (BioTraceTM NT; Pall Corp., FL, United States). The following antibodies were used in this experiment: beta catenin (ab32572; Abcam), inhibin alpha (ab81234; Abcam), HSD17B1 (ab134193; Abcam), MIF (ab227073; Abcam), caspase6 (ab185645; Abcam), laminA/C (MA3-1000; Thermo), and p-laminA/C-S22 (13448; CST, Shanghai, China). The antibodies were diluted to the recommended ratio with Beyotime (P0256; Shanghai, China) diluent. The Western blotting images were analyzed using ImageJ software (National Institutes of Health, Bethesda, MD, United States).
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3

Protein extraction and Western blot analysis

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The GCs were harvested and washed once in PBS, then lysed on ice for 30 min with RIPA buffer (CST, 9806), and supplemented with 1% (v/v) protease inhibitor Cocktail (HY-K0010) and 1% (v/v) phosphatase inhibitors (Cocktail I, HY-K0021; Cocktail II, HY-K0022; and Cocktail III, HY-K0023), which were purchased from MCE (Shanghai, China). Western blotting was performed as described previously (16, 17) . Protein concentrations were determined using a BCA protein assay kit (Transgen Biotech, Beijing, China). Equal amounts of proteins (15-50 µg/lane) were separated by SDS-PAGE (12% acrylamide running gel) and transferred to a nitrocellulose membrane (BioTrace™ NT, Pall Corp, FL, USA). The following antibodies were used in this experiment: beta catenin (ab32572, Abcam), inhibin alpha (ab81234, Abcam), HSD17B1 (ab134193, Abcam), MIF (ab227073, Abcam), caspase6 (ab185645, Abcam), laminA/C (MA3-1000, Thermo), p-laminA/C-S22 (13448, CST, Shanghai, China). The antibodies were diluted to the recommended ratio with Beyotime (P0256, Shanghai, China) diluent. The Western blotting images were analyzed using Image J software (National Institutes of Health, Bethesda, MD, USA).
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