Annexin 5 fl fitc

The standard chemicals, reagents, and the specific antibodies were purchased from Sigma Chemical Co. (St. Louis, MO, USA), Santa Cruz Biotechonolgy (European Support Office, Heidelberg, Germany), Merk & Co (Kenilworth, NJ, USA), Lonza (Basel, Switzerland), R&D Systems (Minneapolis, MN, USA), Qiagen (Hilden, Germany), and from Thermo Fisher Scientific (Waltham, MA, USA). All others reagents were of analytical grade.
For flow cytometry used the following antibodies: Annexin V(FL)-FITC (Santa Cruz, sc-4252 FITC, anti-human, source: E. coli, c = 0.05 µg/µL); PE anti-human CD34 antibody (Biolegend, cod 343606, host species: mouse, c = 0.005 µg/µL); CD133/2(293C3)-APC antibody (Millteny, Order no. 130-090-854, anti-human, clone: 293C3, c = 10 µL/10⁶ cells); human VEGFR2/KDR/Flk-1 Fluorescein-conjugated antibody (R&DSystem, FAB357F, source: monoclonal mouse IgG1 clone # 89106, c = 10 µL/10⁶ cells); Goat F(ab’)2 Anti-Human IgG H&L- (Phycoerythrin) pre-adsorbed antibody (abcam-ab7006, host species: goat, dilution: 1/100); goat anti-human IgG (H+L) cross-adsorbed secondary antibody, FITC (Invitrogen-31531, host species: goat, dilution: 1/100).

Single-cell suspensions of the human iPS-derived cells were treated with 1 × Permeabilization Buffer (e-Bioscience), and then incubated with the primary antibody (anti-VILLIN antibody, (Abcam, ab109516)), followed by the secondary antibody (Goat Anti-Rabbit IgG APC-conjugated Antibody (R&D systems) or Donkey anti-Rabbit IgG (H + L) Secondary Antibody, Alexa Fluor 488 conjugate (Molecular probes)). Single-cell suspensions of the human iPS-derived cells were also incubated with the Ki-67 Antibody (Ki-67) FITC or Annexin V (FL) FITC (both from Santa Cruz Biotechnology). Flow cytometry analysis was performed using a FACS LSR Fortessa flow cytometer (BD Biosciences).