anti-mouse ace2 rabbit polyclonal antibody  (BioChain Institute)

Journal: Molecular Medicine

Article Title: ACE2 Activation Promotes Antithrombotic Activity

doi: 10.2119/molmed.2009.00160

Figure Lengend Snippet: Increased thrombus formation in SHR is associated with lower ACE2 activity in thrombi. (A) FeCl2-induced thrombus weight in SHR and WKY; (B) ACE2 activity in thrombi from SHR and WKY; (C) Representative Western blotting analysis of ACE2 levels. Data were normalized by using β-actin. (D) Quantification of Western blotting data. *P < 0.05 versus WKY (unpaired Student t test, n = 8 to 10). A.U., arbitrary units.

Article Snippet: The membrane was probed with the rabbit polyclonal antibody against ACE2 (sc-20998 [Santa Cruz Biotechnology, Santa Cruz, CA, USA] 1 μg/mL in 1% milk/Tris-buffered saline-Tween solution) overnight at 4oC.

Techniques: Activity Assay, Western Blot

Journal: Molecular Medicine

Article Title: ACE2 Activation Promotes Antithrombotic Activity

doi: 10.2119/molmed.2009.00160

Figure Lengend Snippet: ACE and ACE2 activity in thrombi of SHR and WKY rats. (A) ACE activity in thrombi of SHR and WKY. No significant differences were found between SHR and WKY. (B) Ratio between ACE2/ACE activity in thrombi of SHR and WKY. *P < 0.05 versus WKY (unpaired Student t test, n = 10 to 11). A.U., arbitrary units.

Article Snippet: The membrane was probed with the rabbit polyclonal antibody against ACE2 (sc-20998 [Santa Cruz Biotechnology, Santa Cruz, CA, USA] 1 μg/mL in 1% milk/Tris-buffered saline-Tween solution) overnight at 4oC.

Techniques: Activity Assay

Journal: Molecular Medicine

Article Title: ACE2 Activation Promotes Antithrombotic Activity

doi: 10.2119/molmed.2009.00160

Figure Lengend Snippet: Antithrombotic effect of XNT on rats. FeCl2-induced thrombus weight in WKY and SHR treated with (A) DX600 and (B) XNT. *P < 0.05 (unpaired Student t test). (C) ACE2 activity in thrombi of WKY and SHR treated with either XNT or DX600. *P < 0.05 versus WKY rats, #P < 0.05 versus untreated SHR (one-way ANOVA followed by the Bonferroni posttest). Each column represents mean ± SEM (n = 7 to 9).

Article Snippet: The membrane was probed with the rabbit polyclonal antibody against ACE2 (sc-20998 [Santa Cruz Biotechnology, Santa Cruz, CA, USA] 1 μg/mL in 1% milk/Tris-buffered saline-Tween solution) overnight at 4oC.

Techniques: Activity Assay

Journal: Hypertension

Article Title: Overexpression of Angiotensin-Converting Enzyme 2 in the Rostral Ventrolateral Medulla Causes Long-Term Decrease in Blood Pressure in the Spontaneously Hypertensive Rats

doi: 10.1161/01.hyp.0000259942.38108.20

Figure Lengend Snippet: Figure 1. ACE2 immunoreactivity in the rat medulla oblongata. A, Coronal sections of the medulla oblongata were immunostained with polyclonal anti-ACE2 antibody (sc-20998, Santa Cruz Biotechnology) as described in the Methods section. Sections incubated without primary antibody were used as negative control (a and e). The low-magnification photographs of sections (top) show the distribution of ACE2 immunoreactivity in the caudal part of medulla oblongata including area postrema (AP) and the dorsal motor nucleus of the vagus (b), the NTS (c), and the ventrolateral medulla, including nucleus ambiguus and RVLM (d). The high-magnification photographs of sections (bottom) show ACE2 immunoreactivity of neurons in the AP (f), neurons (arrowhead) and neuropils (arrow) of the NTS (g) and neurons of the RVLM (h). The immunoreactivity in the RVLM neurons was observed in cytoplasm of neurons (arrowhead) and its proxi- mal dendrite (arrow). The scale bar denotes 100 m in the top and 20 m in the bottom. B, Coronal sections of the medulla oblongata were immunostained with polyclonal anti-ACE2 antibody (GTX15348, GeneTex) as described in the Methods section. The low- magnification photographs of sections show negative immunostaining controls consisted of exclusion and preadsorption of the primary antibodies for ACE2 (a and b, respectively). The corresponding low-magnification photograph (c) shows the distribution of ACE2 immunoreactivity in the ventrolateral medulla. The distribution of ACE2 immunoreactivity using this antibody was similar to that of immunoreactivity using the Santa Cruz antibody. The high-magnification photograph shows ACE2 immunoreactivity in the RVLM neurons (d). The scale bar denotes 100 m in a, b, and c, and 20 m in d.

Article Snippet: They were incubated with rabbit polyclonal antibody to ACE2 (1:50, sc-20998, Santa Cruz Biotechnology) or another rabbit polyclonal antibody to ACE2 (1:500, GTX15348, GeneTex) containing 0.3% BSA in PBS containing 0.3% Triton X100 overnight at 4°C followed by incubation with biotinylated goat anti-rabbit IgG for 120 minutes and avidin–biotin–peroxidase complex reagents for 60 minutes and stained with diaminobenzidine solution for 8 minutes according to the manufacturer’s instructions (Vector Laboratories).

Techniques: Incubation, Negative Control, Immunostaining

Journal: Hypertension

Article Title: Overexpression of Angiotensin-Converting Enzyme 2 in the Rostral Ventrolateral Medulla Causes Long-Term Decrease in Blood Pressure in the Spontaneously Hypertensive Rats

doi: 10.1161/01.hyp.0000259942.38108.20

Figure Lengend Snippet: Figure 2. ACE2 protein levels in the RVLM of WKY rats and SHRs. A, Representative autoradiogram of ACE2 protein levels in the RVLM: Western blot analysis was used to measure ACE2 levels from 20 g of total cell lysate isolated from the RVLM punches as described in the Methods section. Data were nor- malized using -tubulin. B, Quantitation of the ACE2 protein band. *P0.05 vs WKY rats. Data were meanSEM (n4 in each strain).

Article Snippet: They were incubated with rabbit polyclonal antibody to ACE2 (1:50, sc-20998, Santa Cruz Biotechnology) or another rabbit polyclonal antibody to ACE2 (1:500, GTX15348, GeneTex) containing 0.3% BSA in PBS containing 0.3% Triton X100 overnight at 4°C followed by incubation with biotinylated goat anti-rabbit IgG for 120 minutes and avidin–biotin–peroxidase complex reagents for 60 minutes and stained with diaminobenzidine solution for 8 minutes according to the manufacturer’s instructions (Vector Laboratories).

Techniques: Western Blot, Isolation, Quantitation Assay

Journal: Hypertension

Article Title: Overexpression of Angiotensin-Converting Enzyme 2 in the Rostral Ventrolateral Medulla Causes Long-Term Decrease in Blood Pressure in the Spontaneously Hypertensive Rats

doi: 10.1161/01.hyp.0000259942.38108.20

Figure Lengend Snippet: Figure 3. Transduction of the SHR RVLM with lenti-ACE2. After termination of the experiment, lentiviral-injected rats were used to eval- uate transgene expression by GFP fluorescence and ACE2 immunostaining as described in the Methods section. A, Representative photograph of lenti-ACE2–injected medulla oblongata: the GFP expression was restricted to the bilateral RVLM. The scale bar denotes 1 mm at the top and 50 m at the bottom. B, Overexpression of ACE2: low-magnification photographs show the lenti-ACE2–injected RVLM. ACE2 immunoreactivity was stronger in the lenti-ACE2–injected site (right) compared with its endogenous expression in the neu- rons (left). The scale bar denotes 200 m at the top and 50 m at the bottom. C, Colocalization of GFP and ACE2 in the lenti-ACE2– injected RVLM: GFP was colocalized with ACE2 in most of the cells. Arrows identify colocalization in individual cells, whereas areas of cells clusters representing both GFP and ACE2 are marked with asterisks. The scale bars denote 200 m.

Article Snippet: They were incubated with rabbit polyclonal antibody to ACE2 (1:50, sc-20998, Santa Cruz Biotechnology) or another rabbit polyclonal antibody to ACE2 (1:500, GTX15348, GeneTex) containing 0.3% BSA in PBS containing 0.3% Triton X100 overnight at 4°C followed by incubation with biotinylated goat anti-rabbit IgG for 120 minutes and avidin–biotin–peroxidase complex reagents for 60 minutes and stained with diaminobenzidine solution for 8 minutes according to the manufacturer’s instructions (Vector Laboratories).

Techniques: Transduction, Injection, Expressing, Immunostaining, Over Expression

Journal: Hypertension

Article Title: Overexpression of Angiotensin-Converting Enzyme 2 in the Rostral Ventrolateral Medulla Causes Long-Term Decrease in Blood Pressure in the Spontaneously Hypertensive Rats

doi: 10.1161/01.hyp.0000259942.38108.20

Figure Lengend Snippet: Figure 4. ACE2 protein levels in the RVLM after ACE2 gene transfer. A, Representative autoradiogram of ACE2 protein levels in the RVLM: Western blot analysis was used to measure ACE2 protein levels from the RVLM punches of noninjected WKY rats and SHRs and SHRs 6 weeks after lenti-ACE2 injection. Data were normalized using -tubulin. B, Quantitation of the ACE2 protein band. Data were meanSEM (n4 in each group).

Article Snippet: They were incubated with rabbit polyclonal antibody to ACE2 (1:50, sc-20998, Santa Cruz Biotechnology) or another rabbit polyclonal antibody to ACE2 (1:500, GTX15348, GeneTex) containing 0.3% BSA in PBS containing 0.3% Triton X100 overnight at 4°C followed by incubation with biotinylated goat anti-rabbit IgG for 120 minutes and avidin–biotin–peroxidase complex reagents for 60 minutes and stained with diaminobenzidine solution for 8 minutes according to the manufacturer’s instructions (Vector Laboratories).

Techniques: Western Blot, Injection, Quantitation Assay

Journal: Hypertension

Article Title: Overexpression of Angiotensin-Converting Enzyme 2 in the Rostral Ventrolateral Medulla Causes Long-Term Decrease in Blood Pressure in the Spontaneously Hypertensive Rats

doi: 10.1161/01.hyp.0000259942.38108.20

Figure Lengend Snippet: Figure 5. Effect of lenti-ACE2 on MAP and HR in WKY rats and SHRs. Lenti-GFP or lenti-ACE2 was injected into the bilateral RVLM of WKY rats (A) and SHRs (B), and MAP (left) and HR (right) were recorded in a conscious state using radioteremetry as described in the Methods section. Lenti- ACE2–injected SHRs showed a time- dependent decrease in MAP and HR. Data are meanSEM (n4 per group in WKY rats and n6 per group in SHRs) *P0.05 vs GFP control groups.

Article Snippet: They were incubated with rabbit polyclonal antibody to ACE2 (1:50, sc-20998, Santa Cruz Biotechnology) or another rabbit polyclonal antibody to ACE2 (1:500, GTX15348, GeneTex) containing 0.3% BSA in PBS containing 0.3% Triton X100 overnight at 4°C followed by incubation with biotinylated goat anti-rabbit IgG for 120 minutes and avidin–biotin–peroxidase complex reagents for 60 minutes and stained with diaminobenzidine solution for 8 minutes according to the manufacturer’s instructions (Vector Laboratories).

Techniques: Injection, Control