Proteins from corneal epithelial samples were extracted using the PARIS kit (Thermo Fisher). Total protein concentration was measured using DC protein assay kit (500–0112, Bio-Rad). 20 μg protein from cell lysates was subjected to 7.5% SDS-PAGE and transferred to Hybond ECL nitrocellulose membrane (Amersham Biosciences, Piscataway, NJ). After incubation with appropriate primary and secondary antibodies, the blots were detected with the Supersignal Femto Chemiluminescent Substrates (ThermoFisher, MA, USA). For re-probing, blots were washed in Western blot stripping buffer (Thermo Fisher) for 15 min before proceeding with new blotting. Anti-ACE2 (ab108252, 1:1000) and anti-TMPRSS2 (ab92323, 1:1000) antibodies were purchased from Abcam. β-actin detected by its antibody (#4970, 1:2000, Cell Signaling) was used for loading control. Horseradish peroxidase-tagged secondary anti-rabbit IgG was from Cell Signaling (#7074, 1:2000).
Anti tmprss2 ab92323
Manufactured by Abcam
Anti-TMPRSS2 (ab92323) is a lab equipment product. It is an antibody that binds to the TMPRSS2 protein.
Automatically generated - may contain errors
Lab products found in correlation
Western blot stripping buffer, by Thermo Fisher Scientific (1 mentions)
Paris kit, by Thermo Fisher Scientific (1 mentions)
Supersignal femto chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Anti ace2 ab108252, by Abcam (1 mentions)
Horseradish peroxidase tagged secondary anti rabbit igg, by Cell Signaling Technology (1 mentions)
Dc protein assay kit, by Bio-Rad (1 mentions)
Immobilon p polyvinylidene difluoride pvdf membrane, by Merck Group (1 mentions)
Complete ultra protease inhibitor cocktail, by Roche (1 mentions)
Gtx117370, by GeneTex (1 mentions)
Gtx117425, by GeneTex (1 mentions)
Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions)
2 protocols using anti tmprss2 ab92323
1
Western Blot Analysis of ACE2 and TMPRSS2
2
Western Blot Analysis of Transmembrane Serine Proteases
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Cells were lysed in lysis buffer (1% NP-40, 20 mM Tris-HCl [pH 7.5], 150 mM NaCl, 5 mM EDTA) supplemented with cOmplete ULTRA protease inhibitor cocktail (Roche Diagnostics, Mannheim, Germany). Lysate samples were resolved by SDS-PAGE and transferred onto Immobilon-P polyvinylidene difluoride (PVDF) membranes (Merck, Burlington, MA). Blots were incubated with the following primary antibodies: anti-TMPRSS2 (ab92323; Abcam, Cambridge, UK) antibody diluted with 5% skim milk in TBST (25 mM Tris-HCl [pH 7.5], 137 mM NaCl, 2.7 mM KCl) and anti-TMPRSS11D (GTX117370; GeneTex, Irvine, CA), anti-TMPRSS11E (PA5-48775; Invitrogen, Thermo Fisher Scientific), and anti-TMPRSS13 (GTX117425; GeneTex) antibodies diluted with Signal Booster (Beacle, Kyoto, Japan). Horseradish peroxidase (HRP)-conjugated anti-β-actin antibody (PM053-7; MBL, Nagoya, Japan) was used as a loading control. Immune complexes were detected using HRP-conjugated secondary antibodies and Immobilon Western chemiluminescent HRP substrate (Merck).
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