Alexa fluor 594 lgg antibody

Cells were cultured on coverslips were fixed and then probed with primary antibody, washed with PBST, probed with secondary antibody in the dark and washed with PBST again. The cells were stained with Vectashield mounting medium containing 4′, 6- diamidino-2-phenylindole (DAPI; Vector Laboratories) before observed under a confocal microscope (Olympus FV1000). Primary antibodies used for probing iPS cells were anti-Oct4 (sc-8628, Santa Cruz), anti-SSEA-1 (mab34301, Millipore) and anti-Nanog (sc-33760, Santa Cruz). As for probing EBs, primary antibodies used were anti-alpha smooth muscle Actin (ab5694, Abcam), anti-Gata4 (sc-25310, Santa Cruz) and anti-Nestin (mab2736, R&D). Secondary antibodies used to probe iPS cells were Alexa Fluor 594 lgG antibody (Invitrogen) while Alexa Fluor 488 lgG antibody (Invitrogen) was used to probe EBs.

E14 cells were cultured on coverslips and BrdU at a concentration of 3 μg/ml was added into the medium. The cells were fixed with 4% paraformaldehyde (PFA), washed with PBS. The primary antibody used to probe the cells was anti-BrdU (sc-32323, Santa Cruz) while the secondary antibody used was Alexa Fluor 594 lgG antibody (Invitrogen).