The fat tissues from 6- and 16-week-old rat offspring were collected and incubated in 4% paraformaldehyde, embedded in paraffin, and sectioned. The sections were incubated with anti-Ang II (1∶200, Abcam, UK) for 2 h at 37°C and then with goat anti-rabbit IgG conjugate (Santa Cruz, USA) for 30 min at 37°C. After each incubation, the slides were washed 3 times with Tris-buffered saline with Tween 20 (TBST) for 5 min each. The slides were then counterstained with Mayer's hematoxylin for 15 s. When observed and photographed under the microscope, positive regions appear brown and yellow. Light micrographs were captured with a color video camera (Nikon Microscope E-100, Japan) and analyzed with image analysis software (Image Pro Plus, USA).
Anti ang 2
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti-Ang II is a laboratory reagent used for the detection and quantification of angiotensin II, a peptide hormone involved in the regulation of blood pressure and fluid balance. It is commonly used in research applications to study the role of angiotensin II in various physiological and pathological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti ang 1, by Abcam (2 mentions)
Image pro plus, by Nikon (1 mentions)
Ab124734, by Abcam (1 mentions)
Ab199956, by Abcam (1 mentions)
Chemidoc touch gel imaging system, by Bio-Rad (1 mentions)
Anti enos, by Abcam (1 mentions)
Mouse anti actin, by Beyotime (1 mentions)
Hrp labeled goat anti mouse igg, by Beyotime (1 mentions)
Hrp labeled goat anti rabbit igg, by Beyotime (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Anti neun, by Merck Group (1 mentions)
Beyoecl plus, by Beyotime (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Abcepta (1 mentions)
4 protocols using anti ang 2
1
Immunohistochemical Analysis of Angiotensin II in Rat Adipose Tissue
2
Protein Expression Analysis in Rat Aorta
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Total protein was extracted from the rat’s thoracic aorta with a protein extraction kit (invent, SA-03-BV). Afterward, protein samples were separated by SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) polyvinylidene fluoride (PVDF) electrophoresis, and target protein bands in the gel were transferred to PVDF membrane. Subsequently, the PVDF membrane was blocked with 5% milk for 1 h, and was incubated with primary antibody (anti-eNOS, 1:1000, Abcam, ab199956, Cambridge, MA, USA; anti-AngII, 1:1000, Abcam, ab124734, Cambridge, MA, USA) overnight at 4°C. Finally, the PVDF membrane strip was incubated with horseradish peroxidase (HRP)-conjugated second antibody (Wuhan Sanying, SA00001-2) for 1 h, and was visualized by ChemiDoc™ Touch Gel imaging system (Bio-Rad, Hercules, CA, USA). The expression of target protein was measured using Image-Pro Plus 6.0 software,17 (link)
and was compared with GAPDH.
and was compared with GAPDH.
3
Engeletin-Mediated Angiogenesis Inhibition
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Engeletin (>99.0% pure, molecular weight: 434.39, CAS: 572-31-6) was obtained from SenBeiJia Biological Technology (China) and was dissolved using dimethyl sulfoxide (DMSO) to prepare a stock solution.
Axitinib (a VEGF receptor (R) inhibitor, Cat No. SC0137-25 mg) and anti-VEGF (Cat No. AV202) were purchased from Beyotime Biotechnology (China). Anti-vasohibin-1 antibody (Cat No.ab176114), anti-Ang-2 (Cat No. ab180820), and anti-Ang-1 (Cat No. ab102015) were from Abcam (UK). Anti-p-Tie2 (Cat No. SAB4503999) and anti-NeuN (Cat No. MEB377) were from Sigma (China). Vasohibin-2 (Cat No. 2923051) was obtained from EMD Millipore (Germany). HRP-labeled goat anti-rabbit IgG (Cat No. A0208), HRP-labeled goat anti-mouse IgG (Cat No. A0216), mouse anti-actin (Cat No. AA128), and BeyoECL Plus (Cat No. P0018FS) were from Beyotime Biotechnology. The 4-0 filament (filament diameter: 0.25 mm; tip diameter: 0.34 mm) was purchased from Beijing Shadong Biology Company (China). RPMI-1640 (Cat No. 11875-093) was from Gibco (USA). Fetal bovine serum (FBS; Cat No.13011-8611) was from Sijiqing (China).
Axitinib (a VEGF receptor (R) inhibitor, Cat No. SC0137-25 mg) and anti-VEGF (Cat No. AV202) were purchased from Beyotime Biotechnology (China). Anti-vasohibin-1 antibody (Cat No.ab176114), anti-Ang-2 (Cat No. ab180820), and anti-Ang-1 (Cat No. ab102015) were from Abcam (UK). Anti-p-Tie2 (Cat No. SAB4503999) and anti-NeuN (Cat No. MEB377) were from Sigma (China). Vasohibin-2 (Cat No. 2923051) was obtained from EMD Millipore (Germany). HRP-labeled goat anti-rabbit IgG (Cat No. A0208), HRP-labeled goat anti-mouse IgG (Cat No. A0216), mouse anti-actin (Cat No. AA128), and BeyoECL Plus (Cat No. P0018FS) were from Beyotime Biotechnology. The 4-0 filament (filament diameter: 0.25 mm; tip diameter: 0.34 mm) was purchased from Beijing Shadong Biology Company (China). RPMI-1640 (Cat No. 11875-093) was from Gibco (USA). Fetal bovine serum (FBS; Cat No.13011-8611) was from Sijiqing (China).
4
Immunohistochemical Analysis of Ang-1 and Ang-2 in Post-SAH Brain
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Immunohistochemistry was performed to study Ang-1 and Ang-2 expression in post SAH hippocampuses and cortices. The sections were deparaffinized and rehydrated in graded concentrations of ethanol and finally in distilled water. The sections were incubated in 3% hydrogen peroxide for 5 minutes to block the endogenous peroxidase activity. To retrieve the hidden epitopes of Ang-1 and Ang-2 proteins, the sections were placed in 10-mmol/L citrate buffer (pH, 6.0), heated in a microwave oven at 95˚C for 30 minutes, cooled at 25˚C for 20 minutes, and then rinsed in phosphate-buffered saline (PBS). The nonspecific protein binding was blocked by incubating the sections in 5% horse serum in PBS for 1hour at 25˚C. The sections were then probed with primary antibodies (anti-Ang-1, 1:1,000, Abcam, Cambridge, MA, USA; anti-Ang-2, 1:500, Abcam) overnight at 4˚C, followed by a 15-minute wash in PBS. The sections were then incubated in horseradish peroxidase (HRP)-conjugated secondary antibody (1:500, Abgent) for 60 minutes at 25˚C. Diaminobenzidine was used as a chromogen, and hematoxylin as a counterstain.
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