primary mouse anti-ace monoclonal antibody  (Santa Cruz Biotechnology)

Journal: International journal of molecular sciences

Article Title: Long Chain N3-PUFA Decreases ACE2 Protein Levels and Prevents SARS-CoV-2 Cell Entry.

doi: 10.3390/ijms232213825

Figure Lengend Snippet: Figure 1. DHA reduces both ACE1 and ACE2 levels in key rat tissues. Western blotting was used to measure ACE1 and ACE2 levels relative to total protein load, as

Article Snippet: Mouse anti-ACE1 monoclonal primary antibody (Catalog #: sc-23908, Santa Cruz, Dallas, TX, USA), together Int.

Techniques: Western Blot

Journal: International journal of molecular sciences

Article Title: Long Chain N3-PUFA Decreases ACE2 Protein Levels and Prevents SARS-CoV-2 Cell Entry.

doi: 10.3390/ijms232213825

Figure Lengend Snippet: Figure 2. DHA differentially modulates ACE1 and ACE2 levels in growing and quiescent EA.hy926 en- dothelial cells. Western blotting was used to compare ACE1 and ACE2 relative to total protein load, as measured by Ponceau S in growing cells treated with ALA, EPA, or DHA at the indicated concentrations (µM) for (a) 8 and (b) 24 h, as well as in quiescent cells treated with n3-PUFA for (c) 8 and (d) 24 h. The band intensities of ACE2 and ACE1 were quantified and are graphically presented in panels (e,f), respectively; the 100 kDa band (grey bars) represents non-glycosylated ACE2, whereas the 130 kDa band (black bars) corresponds to N-glycosylated ACE2. Data are presented as mean ± SEM, n = 3; bars not sharing a common letter in the graphs are significantly different (p < 0.05) based on Duncan’s multiple range or LSD post-hoc tests.

Article Snippet: Mouse anti-ACE1 monoclonal primary antibody (Catalog #: sc-23908, Santa Cruz, Dallas, TX, USA), together Int.

Techniques: Western Blot

Journal: International journal of molecular sciences

Article Title: Long Chain N3-PUFA Decreases ACE2 Protein Levels and Prevents SARS-CoV-2 Cell Entry.

doi: 10.3390/ijms232213825

Figure Lengend Snippet: Figure 5. Schematic of proposed mechanism of action. LCn3-PUFAs, EPA and DHA, reduce ACE2 protein levels. These n3-PUFAs may also decrease ACE2 glycosylation. These mechanisms may explain how DHA blocks SARS-CoV-2 pseudovirus entry into HEK293 cells. LCn3-PUFAs also downregulate ACE1 protein levels, thus maintaining the balance between ACE1 and ACE2, and diminishing the risk of adverse CVD outcomes. This figure was prepared by S. Huang using Microsoft PowerPoint software version 16.16.27.

Article Snippet: Mouse anti-ACE1 monoclonal primary antibody (Catalog #: sc-23908, Santa Cruz, Dallas, TX, USA), together Int.

Techniques: Glycoproteomics, Software

Journal: Frontiers in endocrinology

Article Title: SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β -Cells and in the Human Pancreas Microvasculature.

doi: 10.3389/fendo.2020.596898

Figure Lengend Snippet: FIGURE 1 | ACE2 staining pattern in human pancreas. Immunohistochemistry for ACE2 in human pancreatic tissue sections (case #110118) using R&D MAB933 antibody. ACE2 is markedly expressed in microvasculature associated cells (A, B) in some rare ductal cells (C, D) and in a subset of endocrine cells within pancreatic islets (E, F). Scale bars in (A, C, E) 150 µm. Scale bars in (B, D, F) 70 µm. Zoom-in images are reported in (B, D, F).

Article Snippet: For ACE2-CD31 double immunofluorescence staining, FFPE pancreas sections were incubated overnight at +4°C with November 2020 | Volume 11 | Article 596898 primary antibody monoclonal mouse anti-human ACE2 (cat. MAB933, R&D System, Minneapolis, MS, USA) diluted 1:33 in PBS 1× supplemented with 3% BSA and with polyclonal rabbit anti-CD31 (cat. Ab28364, Abcam, Cambridge, UK) diluted 1:50 in the same blocking buffer.

Techniques: Staining, Immunohistochemistry

Journal: Frontiers in endocrinology

Article Title: SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β -Cells and in the Human Pancreas Microvasculature.

doi: 10.3389/fendo.2020.596898

Figure Lengend Snippet: FIGURE 2 | In microvasculature, ACE2 is putatively expressed in pericytes. (A) Representative image of human pancreatic Formalin-Fixed Paraffin Embedded (FFPE) section stained for ACE2 in case #301118. In panel-a, a representative image of a pancreatic section showing two adjacent lobules (blue and red dotted lines) with different staining for ACE2 in endothelial cells/pericytes. A specific segmentation of the two lobules with high (blue) (zoom-in, panel-b) and low or null expression of ACE2 (red) (zoom-in, panel-c) is shown, suggesting lobularity of ACE2 expression in exocrine endothelial cells/pericytes of human pancreas. Scale bar in panel-a: 100 µm. Scale bar in panels-b and -c: 30 µm. (B) Double immunofluorescence staining of ACE2 (green) and CD31 (red) in FFPE pancreas sections from Body01A of Case #110118 (panels-a to -d) and of Body01B of Case #141117 (panels-e to -i). Digital zoom-in overlay images are shown in panels-d, -h and -i. Scale bar in panels-d and -g: 100 mm.

Article Snippet: For ACE2-CD31 double immunofluorescence staining, FFPE pancreas sections were incubated overnight at +4°C with November 2020 | Volume 11 | Article 596898 primary antibody monoclonal mouse anti-human ACE2 (cat. MAB933, R&D System, Minneapolis, MS, USA) diluted 1:33 in PBS 1× supplemented with 3% BSA and with polyclonal rabbit anti-CD31 (cat. Ab28364, Abcam, Cambridge, UK) diluted 1:50 in the same blocking buffer.

Techniques: Staining, Expressing

Journal: Frontiers in endocrinology

Article Title: SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β -Cells and in the Human Pancreas Microvasculature.

doi: 10.3389/fendo.2020.596898

Figure Lengend Snippet: FIGURE 3 | ACE2 immunohistochemistry staining pattern in human pancreatic islets using MAB933, Ab15348, and Ab108252 antibodies. (A) Aligned sequences and structures of recently described ACE2 isoforms, long-ACE2 (805aa, ~110 kDa) and short-ACE2 (459aa, ~50kDa). Main ACE2 protein domains are reported with different colors. (B) R&D MAB933, (C) Abcam Ab15348 and (D) Ab108252 antibody predicted target sequence within the two ACE2 isoforms, alongside with immunohistochemistry staining distribution in pancreatic islets. Scale bars in (B–D) are 100 mm.

Article Snippet: For ACE2-CD31 double immunofluorescence staining, FFPE pancreas sections were incubated overnight at +4°C with November 2020 | Volume 11 | Article 596898 primary antibody monoclonal mouse anti-human ACE2 (cat. MAB933, R&D System, Minneapolis, MS, USA) diluted 1:33 in PBS 1× supplemented with 3% BSA and with polyclonal rabbit anti-CD31 (cat. Ab28364, Abcam, Cambridge, UK) diluted 1:50 in the same blocking buffer.

Techniques: Immunohistochemistry, Staining, Sequencing

Journal: Frontiers in endocrinology

Article Title: SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β -Cells and in the Human Pancreas Microvasculature.

doi: 10.3389/fendo.2020.596898

Figure Lengend Snippet: FIGURE 4 | In human pancreatic islets, ACE2 is preferentially expressed in insulin-producing b-cells. Triple immunofluorescence staining and image analysis of FFPE human pancreatic section stained for insulin (red), glucagon (blue) and ACE2 (green). (A) Representative islets of two different cases. Panels-a to -g: representative pancreatic islet of FFPE pancreas block Body01A of case #110118. Panels-h to -m: representative pancreatic islet of FFPE pancreas block Body01B of case #141117. Panels-e to -g: digital zoom in images of the pancreatic islet shown in panel-d. Panels-l and -m: digital zoom in images of the pancreatic islet shown in panel-k. Scale bar in panels-d and -k: 100 µm. (B) Colocalization rate analysis of overlapping ACE2-insulin and ACE2-glucagon in 128 single pancreatic islets of seven different cases. p-value was calculated using Wilcoxon matched-pairs signed rank test. On the right: colocalization rate analysis of ACE2-insulin and ACE2-glucagon in each of the seven cases analyzed. For each case, a total of 7–11 islets/section were analyzed. (C) Analysis of the intensity of ACE2 islet-related signals in the cases analyzed. Values are shown as fluorescence intensity of each islet reported as the sum of gray-scale values for each pixel normalized for the islets area (ROI, mm2).

Article Snippet: For ACE2-CD31 double immunofluorescence staining, FFPE pancreas sections were incubated overnight at +4°C with November 2020 | Volume 11 | Article 596898 primary antibody monoclonal mouse anti-human ACE2 (cat. MAB933, R&D System, Minneapolis, MS, USA) diluted 1:33 in PBS 1× supplemented with 3% BSA and with polyclonal rabbit anti-CD31 (cat. Ab28364, Abcam, Cambridge, UK) diluted 1:50 in the same blocking buffer.

Techniques: Staining, Blocking Assay

Journal: Frontiers in endocrinology

Article Title: SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β -Cells and in the Human Pancreas Microvasculature.

doi: 10.3389/fendo.2020.596898

Figure Lengend Snippet: FIGURE 5 | A short-ACE2 isoform is prevalently expressed in human b-cell line EndoC-bH1. Western Blot and Immunofluorescence analysis of EndoC-bH1 using (A) R&D monoclonal MAB933, (B) Abcam polyclonal Ab15348 and (C) Abcam monoclonal Ab108252 anti-ACE2 antibody. For each antibody adopted, specific target sequence is reported within the aligned ACE2 isoforms. In western blot analysis, molecular weight markers (from 15 kDa to 250 kDa) are reported; red arrows indicate long-ACE2 isoform (expected band of ~110kDa), while blue arrows indicate short-ACE2 isoform (~50kDa). (D) ACE2 (R&D MAB933) and insulin double immunofluorescence analysis in EndoC- bH1 cultured cells. Negative isotype primary antibody control (relative to ACE2 primary antibody) is shown in panel-a. Insulin (red) and ACE2 (green) are reported in panels-b and -c, while overlay is reported in panel-d. Digital zoom-in images are reported from panels-e to -h. Scale bar in panel-h = 15 µm.

Article Snippet: For ACE2-CD31 double immunofluorescence staining, FFPE pancreas sections were incubated overnight at +4°C with November 2020 | Volume 11 | Article 596898 primary antibody monoclonal mouse anti-human ACE2 (cat. MAB933, R&D System, Minneapolis, MS, USA) diluted 1:33 in PBS 1× supplemented with 3% BSA and with polyclonal rabbit anti-CD31 (cat. Ab28364, Abcam, Cambridge, UK) diluted 1:50 in the same blocking buffer.

Techniques: Western Blot, Sequencing, Molecular Weight, Cell Culture, Control

Journal: Frontiers in endocrinology

Article Title: SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β -Cells and in the Human Pancreas Microvasculature.

doi: 10.3389/fendo.2020.596898

Figure Lengend Snippet: FIGURE 6 | qRT-Real Time PCR analysis of ACE2 mRNA. (A–D) ACE2 raw Ct values results in lung tissue (n = 1, in duplicate), in enzymatic-isolated human pancreatic islets samples (n = 4), in LCM-microdissected islets (n = 5) and in EndoC-bH1 (n = 4). (E–H) ACE2 expression values normalized using GAPDH and b2- microglobulin of the samples analysed in A–D. Values are reported as 2−dCt. Mean ± S.D. values are shown.

Article Snippet: For ACE2-CD31 double immunofluorescence staining, FFPE pancreas sections were incubated overnight at +4°C with November 2020 | Volume 11 | Article 596898 primary antibody monoclonal mouse anti-human ACE2 (cat. MAB933, R&D System, Minneapolis, MS, USA) diluted 1:33 in PBS 1× supplemented with 3% BSA and with polyclonal rabbit anti-CD31 (cat. Ab28364, Abcam, Cambridge, UK) diluted 1:50 in the same blocking buffer.

Techniques: Real-time Polymerase Chain Reaction, Isolation, Expressing

Journal: Frontiers in endocrinology

Article Title: SARS-CoV-2 Receptor Angiotensin I-Converting Enzyme Type 2 (ACE2) Is Expressed in Human Pancreatic β -Cells and in the Human Pancreas Microvasculature.

doi: 10.3389/fendo.2020.596898

Figure Lengend Snippet: FIGURE 7 | ACE2 expression is increased by inflammatory stress. ACE2 mRNA RT-Real-Time PCR analysis in EndoC-bH1 treated or not (CTR) with Palmitate (2.0 mM) (A) or with cytokines (IL-1b + IFNg + TNFa) (Cyt. Mix) (B) for 24 h. Results are reported as mean ± S.D of 2−dCT normalized values. p-values were calculated using Wilcoxon matched-pairs signed rank test. (C) Immunofluorescence analysis of insulin (red, panels-a and -g) and ACE2 (green, panels-b and -h) in EndoC-bH1 not-treated (panels-a to -f) or treated (panels-g to -l) with cytokines for 24 h. Digital zoom-in images are reported in panels-d to -f and in panels-j to -l. Scale bar in panel-f: 10 µm. Scale bar in panel-l: 15 µm. (D) Representative images of ACE2 staining (green) and analysis using micro-confocal High-content screening in EndoC-bH1 treated or not with cytokines (IL-1b + IFNg+TNFa). Panels-a and -b: ACE2 signal (green) and automated cell cytoplasm segmentation and identification. Panels-c and -d: identification of ACE2 granular spots within segmented cytoplasm in panels-a and -b. Each ACE2 granular spot intensity was measured and analyzed. (E) Mean intensity imaging analysis related to data reported in (C) of EndoC-bH1 treated or not with cytokines. Data are reported as individual values alongside with mean ± S.D of RGB gray-intensity measures of 6–11 different experimental points related to two different independent experiments. Individual values alongside with mean ± S.D are reported. P-value was calculated using Mann–Whitney U test. (F) High content screening analysis of Corrected Median Spot intensity of ACE2 signal in EndoC-bH1 treated or not with cytokines. Median intensity values of six different experimental points are reported. P-value was calculated using Mann–Whitney U test (p < 0.05).

Article Snippet: For ACE2-CD31 double immunofluorescence staining, FFPE pancreas sections were incubated overnight at +4°C with November 2020 | Volume 11 | Article 596898 primary antibody monoclonal mouse anti-human ACE2 (cat. MAB933, R&D System, Minneapolis, MS, USA) diluted 1:33 in PBS 1× supplemented with 3% BSA and with polyclonal rabbit anti-CD31 (cat. Ab28364, Abcam, Cambridge, UK) diluted 1:50 in the same blocking buffer.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Staining, High Content Screening, Imaging, MANN-WHITNEY