mouse anti catd Search Results


mouse monoclonal anti-catd antibody  (Becton Dickinson)
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Becton Dickinson mouse monoclonal anti-catd antibody
Sorting of non-Cab45 cargos is not affected in Cab45 phosphomutants (related to ). (A) Western blot analysis of the secretion of <t>CatD</t> in HeLa control and Fam20C-KO cells. Endogenous CatD was trapped in the Golgi at 20°C and released for 1 h at 37°C. The supernatants (upper panel) and cell lysates (lower panels) were tested for CatD by Western blotting. β-Actin was used as loading control. (B) Representative immunofluorescence images of colocalization of vesicular SBP-EGFP-CatD with the lysosomal <t>marker</t> <t>Lamp1.</t> HeLa cells were transfected with RUSH construct and incubated for 40 min with biotin. Cells were fixed and stained with α-Lamp1 antibody. Scale bar, 10 µm. The magnification of the inset is shown in the lower right pane (scale bar, 1 µm). (C) Representative immunofluorescence images of RUSH experiments showing OPN transport in HeLa control, Cab45-KO, Cab45-WT, and Cab45 phosphomutant cell lines. Cells were transfected with OPN-SBP-EGFP and fixed at 0, 20, 40, and 50 min after the addition of biotin. Z-stack images (d = 0.35 µm) were analyzed. The arrowheads indicate cytoplasmic vesicles. Scale bars, 10 µm. (D) Representative immunofluorescence images of RUSH experiments showing cathepsin D (CatD) transport in Cab45-WT and phosphomutant cell lines. Cells were transfected with SBP-EGFP-CatD and fixed at 0, 20, 40, and 60 min after the addition of biotin. Z-stack images (d = 0.35 µm) were analyzed. The arrowheads indicate cytoplasmic vesicles. Scale bars, 10 µm. (E) The numbers of CatD budding vesicles (D) were quantified. The cytoplasmic vesicles were counted at each time point by analyzing z-stack images (d = 0.35 µm). Scatter dot plot represents the means ± SD of at least three independent experiments ( n > 30 cells per condition). Statistical test, Kruskal–Wallis.
Mouse Monoclonal Anti Catd Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Sorting of non-Cab45 cargos is not affected in Cab45 phosphomutants (related to ). (A) Western blot analysis of the secretion of CatD in HeLa control and Fam20C-KO cells. Endogenous CatD was trapped in the Golgi at 20°C and released for 1 h at 37°C. The supernatants (upper panel) and cell lysates (lower panels) were tested for CatD by Western blotting. β-Actin was used as loading control. (B) Representative immunofluorescence images of colocalization of vesicular SBP-EGFP-CatD with the lysosomal marker Lamp1. HeLa cells were transfected with RUSH construct and incubated for 40 min with biotin. Cells were fixed and stained with α-Lamp1 antibody. Scale bar, 10 µm. The magnification of the inset is shown in the lower right pane (scale bar, 1 µm). (C) Representative immunofluorescence images of RUSH experiments showing OPN transport in HeLa control, Cab45-KO, Cab45-WT, and Cab45 phosphomutant cell lines. Cells were transfected with OPN-SBP-EGFP and fixed at 0, 20, 40, and 50 min after the addition of biotin. Z-stack images (d = 0.35 µm) were analyzed. The arrowheads indicate cytoplasmic vesicles. Scale bars, 10 µm. (D) Representative immunofluorescence images of RUSH experiments showing cathepsin D (CatD) transport in Cab45-WT and phosphomutant cell lines. Cells were transfected with SBP-EGFP-CatD and fixed at 0, 20, 40, and 60 min after the addition of biotin. Z-stack images (d = 0.35 µm) were analyzed. The arrowheads indicate cytoplasmic vesicles. Scale bars, 10 µm. (E) The numbers of CatD budding vesicles (D) were quantified. The cytoplasmic vesicles were counted at each time point by analyzing z-stack images (d = 0.35 µm). Scatter dot plot represents the means ± SD of at least three independent experiments ( n > 30 cells per condition). Statistical test, Kruskal–Wallis.

Journal: The Journal of Cell Biology

Article Title: Fam20C regulates protein secretion by Cab45 phosphorylation

doi: 10.1083/jcb.201910089

Figure Lengend Snippet: Sorting of non-Cab45 cargos is not affected in Cab45 phosphomutants (related to ). (A) Western blot analysis of the secretion of CatD in HeLa control and Fam20C-KO cells. Endogenous CatD was trapped in the Golgi at 20°C and released for 1 h at 37°C. The supernatants (upper panel) and cell lysates (lower panels) were tested for CatD by Western blotting. β-Actin was used as loading control. (B) Representative immunofluorescence images of colocalization of vesicular SBP-EGFP-CatD with the lysosomal marker Lamp1. HeLa cells were transfected with RUSH construct and incubated for 40 min with biotin. Cells were fixed and stained with α-Lamp1 antibody. Scale bar, 10 µm. The magnification of the inset is shown in the lower right pane (scale bar, 1 µm). (C) Representative immunofluorescence images of RUSH experiments showing OPN transport in HeLa control, Cab45-KO, Cab45-WT, and Cab45 phosphomutant cell lines. Cells were transfected with OPN-SBP-EGFP and fixed at 0, 20, 40, and 50 min after the addition of biotin. Z-stack images (d = 0.35 µm) were analyzed. The arrowheads indicate cytoplasmic vesicles. Scale bars, 10 µm. (D) Representative immunofluorescence images of RUSH experiments showing cathepsin D (CatD) transport in Cab45-WT and phosphomutant cell lines. Cells were transfected with SBP-EGFP-CatD and fixed at 0, 20, 40, and 60 min after the addition of biotin. Z-stack images (d = 0.35 µm) were analyzed. The arrowheads indicate cytoplasmic vesicles. Scale bars, 10 µm. (E) The numbers of CatD budding vesicles (D) were quantified. The cytoplasmic vesicles were counted at each time point by analyzing z-stack images (d = 0.35 µm). Scatter dot plot represents the means ± SD of at least three independent experiments ( n > 30 cells per condition). Statistical test, Kruskal–Wallis.

Article Snippet: The antisera used were rat monoclonal anti-HA (Roche; catalog #11867423001), rabbit monoclonal anti-KIAA0310 (Thermo Fisher Scientific; catalog #A300-648A-M), mouse monoclonal anti-cis-Golgi matrix protein 130 (BD Biosciences; catalog #610823), mouse monoclonal anti-CNX (Sigma-Aldrich; catalog #C7617), mouse monoclonal anti-p230 (BD Biosciences; catalog #611281), sheep polyclonal anti-TGN46 (AbD Serotec; catalog #AHP500G), mouse monoclonal anti-β-actin (Sigma-Aldrich; catalog #A5441), mouse monoclonal ANTI-FLAG M2-peroxidase (HRP; Sigma-Aldrich; catalog #A8592), mouse monoclonal anti-EEA1 antibody (BD Biosciences; catalog #610457), rabbit polyclonal anti-Rab11 antibody (gift from the De Camilli laboratory, Yale School of Medicine, New Haven, CT, self-made), rabbit monoclonal anti-Lamp1 antibody (Cell Signaling; catalog #9091S), and mouse monoclonal anti-CatD antibody (BD Biosciences; catalog #610801).

Techniques: Western Blot, Immunofluorescence, Marker, Transfection, Construct, Incubation, Staining