Immunohistochemistry (IHC) of whole-mount samples or tissue sections was performed as previously described79 (link). At P6, the primary monoclonal antibodies used were anti-RhoA (21017; NewEast Biosciences, King of Prussia, PA), hamster anti-CD31 (2H8; Chemicon, Temecula, CA), Secondary antibodies used were Alexa 488 fluorescence-conjugated IgGs (Molecular Probes) or Cy3/Cy5 DyLight549/DyeLight649-conjugated IgGs (Jackson ImmunoResearch, West Grove, PA). At P9, blood vessels were visualized using biotinylated isolectin B4 (IB4) (Sigma), followed by fluorescent streptavidin conjugates (Molecular Probes).
Fluorescent streptavidin conjugates
Manufactured by Thermo Fisher Scientific
Fluorescent streptavidin conjugates are protein complexes that bind strongly to the vitamin biotin. They are used in various bioanalytical techniques to detect and visualize biotinylated molecules, such as proteins, nucleic acids, or cell surface receptors.
Automatically generated - may contain errors
Lab products found in correlation
Biotinylated isolectin b4 ib4, by Merck Group (1 mentions)
Cy3 cy5 dylight549 dylight649 conjugated iggs, by Jackson ImmunoResearch (1 mentions)
Alexa fluor 488 conjugated anti gfp a21311, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Biotinylated isolectin b4, by Vector Laboratories (1 mentions)
Anti vegf sc 152, by Santa Cruz Biotechnology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
2 protocols using fluorescent streptavidin conjugates
1
Immunohistochemistry of Vascular Tissues
2
Multicolor Immunohistochemistry of Tissues
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Immunohistochemistry of whole-mount samples or tissue sections was performed as previously described (Kubota et al., 2009 (link)). The primary monoclonal antibodies used were hamster anti-CD31 (MAB1398Z; EMD Millipore), PDGFRα (14-1401; eBioscience), vascular endothelial cadherin (550548; BD), and F4/80 (MCA497R; Serotec). The primary polyclonal antibodies used were Alexa Fluor 488–conjugated anti-GFP (A21311; Molecular Probes), collagen IV (LSL-LB-1407; Cosmo Bio), anti-VEGF (sc-152; Santa Cruz Biotechnology, Inc.), and cleaved caspase-3 (9664; Cell Signaling Technology). The secondary antibodies used were Alexa Fluor 488 fluorescence–conjugated IgGs (Molecular Probes) or Cy3/Cy5 DyLight 549/DyLight 649–conjugated IgGs (Jackson ImmunoResearch Laboratories, Inc.). For nuclear staining, specimens were treated with DAPI (Molecular Probes). In some experiments, blood vessels and monocyte lineage cells were simultaneously visualized using biotinylated isolectin B4 (B-1205; Vector Laboratories) followed by fluorescent streptavidin conjugates (Molecular Probes).
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