anti rabbit goat polyclonal antibody  (Bio-Rad)

Journal: Investigative Ophthalmology & Visual Science

Article Title: Anti-VEGF Agents Clearance Through the Aqueous Outflow Pathway in a Rat Model

doi: 10.1167/iovs.66.6.1

Figure Lengend Snippet: Clearance of anti-VEGF agents. Clearance was evaluated by immunostaining signals in the test groups throughout the timeline of the studies. Immunoreactivity gradually decreased until the immunofluorescent signal was completely negligible after 48 hours. The bevacizumab sections are presented in horizontal ascending order from 3 hours to 48 hours; the top row is at low magnification, and the bottom row is at high magnification (× 100). Mouse anti-rat CD31 ( red ) and DAPI ( blue ) were used as markers for anatomic orientation and detection of Schlemm canal (SC) endothelial cells. Alexa Fluor 488 conjugated donkey anti-human IgG antibody ( green ) was used to detect bevacizumab immunoreactivity. The aflibercept and ranibizumab sections are presented in vertical ascending order from time zero to 48 hours. Mouse monoclonal anti-αSMA ( green ) and DAPI ( blue ) were used for anatomic orientation. Adjacent to the immunostained sections are corresponding sections stained with H&E to assess tissue morphology. Aflibercept was detected by rabbit polyclonal anti-human IgG (269A-14 red, “Rb IgG” or “IgG”) and donkey anti-rabbit Cy3: excitation = 554 nm and emission = 568 nm. Ranibizumab was detected by goat polyclonal anti-human IgG-Fab (A80-114A, red, “G Fab”) and donkey anti-goat Cy3: excitation = 554 nm and emission = 568 nm; rabbit polyclonal anti-human IgG (H/L; #STAR195, pale-blue, “Rb IgG” or “IgG”) and donkey anti-rabbit Cy5: excitation = 649 nm and emission = 667 nm. pH6, high conc., magnification × 20, Zoom 1.

Article Snippet: The primary and secondary antibodies for anti-VEGF agents were as follows : bevacizumab – primary antibody: human IgG (709-545-149; Jackson ImmunoResearch, West Grove, PA, USA); secondary antibody: donkey anti-human IgG labeled with Alexa Fluor 488 (Jackson ImmunoResearch); aflibercept – primary antibody: rabbit polyclonal anti-human IgG (269A, 711-165-152; Cell Marque); secondary antibody: donkey anti-rabbit Cy3: excitation 554 nm, emission 568 nm (Jackson ImmunoResearch); ranibizumab – primary antibodies: goat polyclonal anti-human IgG-Fab (A80-114A; Fortis Life Sciences, Waltham, MA, USA) and rabbit polyclonal anti-human IgG (H/L) (STAR195; Bio-Rad, Netiv HaOr, Haifa, Israel); secondary antibodies: donkey anti-goat Cy3, excitation 554 nm, emission 568 nm, and donkey anti-rabbit Cy5: excitation 649 nm, emission 667 nm (Jackson ImmunoResearch).

Techniques: Immunostaining, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Anti-VEGF Agents Clearance Through the Aqueous Outflow Pathway in a Rat Model

doi: 10.1167/iovs.66.6.1

Figure Lengend Snippet: Negative and positive control groups. Negative control samples were processed with secondary antibodies alone. As expected, no immunofluorescent signal was seen. The only fluorescent staining is of DAPI blue, which represents nuclei in the tissue. Bevacizumab (A2), aflibercept, and ranibizumab are presented in the middle. Magnification × 20, Zoom 1. Positive control samples were processed with all primary and secondary antibodies. Magnification × 20, Zoom 1. A fluorescent signal is seen from all types of antibodies. Bevacizumab (A1, B3) detection in green (anti-human IgG antibody) and endothelial cells in red (anti CD31); corresponding H&E (B1) areas with illustration (B2) of trabecular meshwork (TM), Schlemm’s canal (SC) and episcleral veins in the rat’s aqueous drainage tract. Aflibercept and ranibizumab: smooth muscle in green (mouse monoclonal anti-αSMA, abbreviated αSMA), aflibercept in red (rabbit polyclonal anti-human IgG, abbreviated Rb); ranibizumab in red (goat polyclonal anti-human IgG-Fab, abbreviated G Fab), and pale blue (rabbit polyclonal anti-human IgG [H/L], “Rb IgG”). BF = brightfield.

Article Snippet: The primary and secondary antibodies for anti-VEGF agents were as follows : bevacizumab – primary antibody: human IgG (709-545-149; Jackson ImmunoResearch, West Grove, PA, USA); secondary antibody: donkey anti-human IgG labeled with Alexa Fluor 488 (Jackson ImmunoResearch); aflibercept – primary antibody: rabbit polyclonal anti-human IgG (269A, 711-165-152; Cell Marque); secondary antibody: donkey anti-rabbit Cy3: excitation 554 nm, emission 568 nm (Jackson ImmunoResearch); ranibizumab – primary antibodies: goat polyclonal anti-human IgG-Fab (A80-114A; Fortis Life Sciences, Waltham, MA, USA) and rabbit polyclonal anti-human IgG (H/L) (STAR195; Bio-Rad, Netiv HaOr, Haifa, Israel); secondary antibodies: donkey anti-goat Cy3, excitation 554 nm, emission 568 nm, and donkey anti-rabbit Cy5: excitation 649 nm, emission 667 nm (Jackson ImmunoResearch).

Techniques: Positive Control, Negative Control, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Anti-VEGF Agents Clearance Through the Aqueous Outflow Pathway in a Rat Model

doi: 10.1167/iovs.66.6.1

Figure Lengend Snippet: Three-dimensional volumetric reconstruction. Images of aflibercept and ranibizumab in brightfield, with adjacent side views (SVs) to gain depth perspective of the immunofluorescent signal which gradually diminished over time, from 0 to 48 hours. By 48 hours, the signal is negligible. Immunostaining: smooth muscle in green (mouse monoclonal anti-αSMA, “αSMA”), aflibercept in red (rabbit polyclonal anti-human IgG [269A-14], “Rb IgG”), ranibizumab in red (goat polyclonal anti-human IgG-Fab [A80-114A], “G Fab”) and in pale blue (rabbit polyclonal anti-human IgG [H/L] (#STAR195, “Rb IgG”); SV in black corresponds to aflibercept “Rb IgG”; SV in red corresponds to ranibizumab “G Fab” antibody; SV in pale blue corresponds to ranibizumab “Rb IgG” antibody. pH6; high conc, magnification × 20, Zoom 1.

Article Snippet: The primary and secondary antibodies for anti-VEGF agents were as follows : bevacizumab – primary antibody: human IgG (709-545-149; Jackson ImmunoResearch, West Grove, PA, USA); secondary antibody: donkey anti-human IgG labeled with Alexa Fluor 488 (Jackson ImmunoResearch); aflibercept – primary antibody: rabbit polyclonal anti-human IgG (269A, 711-165-152; Cell Marque); secondary antibody: donkey anti-rabbit Cy3: excitation 554 nm, emission 568 nm (Jackson ImmunoResearch); ranibizumab – primary antibodies: goat polyclonal anti-human IgG-Fab (A80-114A; Fortis Life Sciences, Waltham, MA, USA) and rabbit polyclonal anti-human IgG (H/L) (STAR195; Bio-Rad, Netiv HaOr, Haifa, Israel); secondary antibodies: donkey anti-goat Cy3, excitation 554 nm, emission 568 nm, and donkey anti-rabbit Cy5: excitation 649 nm, emission 667 nm (Jackson ImmunoResearch).

Techniques: Immunostaining