Alexa fluor 633 conjugated donkey anti goat igg

The primary antibodies used were obtained from the following companies: anti-alpha-fetoprotein Ab-2 (1:100 for immunohistochemistry, 1:1000 for Western blotting; RB-365) from Thermo Fisher Scientific, anti-EGF receptor (1:1000, 610017) from BD Biosciences, anti-actin (1:10000, A2066) and anti-phospho-ERK1/2 (1:1000, 05-797R) from Sigma-Aldrich, anti-ERK1/2 (1:1000, 9102) from Cell Signaling Technology, anti-VEGF Ab-1 (1:1000, RB-222) from NeoMarkers, and anti-PECAM-1 (1:250, sc-1506) from Santa Cruz Biotechnology. The secondary antibodies used were Cy3-conjugated goat anti-rabbit IgG (1:500, 111-165-003) from Jackson ImmunoResearch, Alexa Fluor 633–conjugated donkey anti-goat IgG (1:1000, A21082) from Invitrogen, Cy3-labeled goat anti-mouse IgG (1:500, 115-165-003) from Jackson ImmunoResearch, HRP-conjugated goat anti-mouse IgG (1:1000, AP124P) from Millipore, HRP-conjugated goat anti-rabbit IgG (1:1000, 111-035-144) from Jackson ImmunoResearch, and HRP-conjugated goat anti-rat IgG (1:1000) from Chemicon.

Uranyl acetate (UA) dihydrate (purity >98.0%) was purchased from Fluka (Buchs, Switzerland). Collagenase type II was from Worthington Biochemical Corp. (Lakewood, NJ). Percoll was purchased from GE Healthcare UK, Ltd. (Little Chalfont, Buckinghamshire, UK). Trypan blue solution, propidium iodide, Hoechst 33342, and pyronin Y were purchased from Sigma‐Aldrich Co. (St. Louis, MO). Hank's balanced salt solution (HBSS) was from Invitrogen (Carlsbad, CA). Can Get Signal^® solution B was from Toyobo Life Science Department (Osaka, Japan). Citrate buffer solution was from Mitsubishi Chemical Medience (Tokyo, Japan). Histofine Antigen Retrieval Solution pH9^® and Histofine MAX PO kit were from Nichirei Bioscience (Tokyo, Japan). ApopTag^® Plus In Situ Apoptosis Detection Kit was from Chemicon‐Millipore (Temecula, CA). Picrosirius red stain kit was from Polysciences, Inc. (Warrington, PA). The antibodies listed in Table 1 were used as primary antibodies. Alexa Fluor^® 633‐conjugated donkey anti‐goat IgG (Invitrogen), Alexa Fluor^® 546‐conjugated goat anti‐rabbit IgG (Invitrogen), Alexa Fluor 488‐conjugated donkey anti‐mouse IgG (Invitrogen), and Histofine Simple Stain Max PO (Nichirei Bioscience) were used as secondary antibodies.