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2 protocols using ab15154

1

Molecular Mechanisms of Estrogen-Mediated Autophagy

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Estradiol (E2758, purity ≥ 98%), MPP dihydrochloride hydrate (M7068, purity ≥ 97%), PHTPP (SML1355, purity ≥ 98%) and 3-Methyladenine (3-MA, M9281, purity ≥ 99%) were purchased from Sigma (Saint Louis, USA). Antibody for NLRP3 (19771-1-AP) and IL-18 (10663-1-AP) were purchased from Proteintech (Wuhan, China). Antibody for caspase-1 (48847) was purchased from SAB signalway antibody (Maryland, USA). Antibody for GSDMD (A18281) and IL-1β (A16288) were purchased from ABclonal (Wuhan, China). Antibody for Beclin 1 (ab210498), LC3B (ab192890), SQSTM1 (ab56416), Hcy (ab15154), estrogen receptor alpha (ERα) (ab32063), estrogen receptor beta (ERβ) (ab3576), GSDMD (ab219800), and β-actin (ab8226) were purchased from Abcam (Cambridge, UK). FITC - goat Anti-mouse IgG (ab6785), TRITC - goat anti-rabbit IgG (BS10250), FITC - goat anti-rabbit IgG (BS10950), HRP - goat anti-rabbit IgG (BS13278), and HRP - goat anti-mouse IgG (BS12478) were purchased from Bioworld (Bloomington, USA). The prestained protein MW marker (26617) were purchased from Thermo scientific. Control siRNA and ERα siRNA (sc-29305) were purchased from Santa Cruz (Santa Cruz, USA).
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2

Protein Interactions in Situ Quantification

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The proximity ligation assay (Duolink in situ PLATM reagents, Olink Bioscience, Eurogentec, Angers, France) was used to depict and quantify protein interactions in situ as previously described. 15 (link) Interactions were assessed between homocysteine (Ab15154; Abcam) and vinculin (14-9777-82; Thermo Fisher Scientific), SM22α (AF7886; R&D Systems), N-cadherin (GC4; Sigma-Aldrich), and Fibrillin (MAB1919; Sigma-Aldrich). Interactions with normal binding partners were assessed between N-cadherin and β-catenin (AF1329; R&D Systems), and between α-SMA (Ab5694, Abcam) and both SM22α and vinculin.
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