BD PharmingenTM PE Mouse anti-Human Pax-6

Description

Intracellular staining of Pax-6 in neural induction of human embryonic stem ES cells. H9 human ES cells WiCell, Madison, WI were cultured in mTeSR Stem Cell Technologies on plates coated with BD Matrigel hESC-qualified Matrix Cat. No. 354277 . Embryoid bodies EB were made and cultured in medium containing Knockout Serum Replacement Life Technologies without bFGF for 24 hours and then in medium containing 250 ng/ml human recombinant noggin R&D Systems and 10 mu M SB 431542 Tocris for 4 more days. The EB were then plated on BD Matrigel-coated plates and grown in medium with ITS supplement Sigma-Aldrich , noggin, and SB 431542. After growth for 7 days, the cells were collected, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 . The cells were then stained with PE Mouse anti-Human Pax-6 and PerCP-Cy5.5 Mouse anti-Human Sox1 Cat. No. 561549 . The plot was derived from gated events based on light scattering characteristics for the neural induction. Flow cytometry was performed on a BD LSR II flow cytometry system. Intracellular staining of Pax-6 in neural induction of human embryonic stem ES cells. H9 human ES cells WiCell, Madison, WI were cultured in mTeSR Stem Cell Technologies on plates coated with BD Matrigel hESC-qualified Matrix Cat. No. 354277 . Embryoid bodies EB were made and cultured in medium containing Knockout Serum Replacement Life Technologies without bFGF for 24 hours and then in medium containing 250 ng/ml human recombinant noggin R&D Systems and 10 mu M SB 431542 Tocris for 4 more days. The EB were then plated on BD Matrigel-coated plates and grown in medium with ITS supplement Sigma-Aldrich , noggin, and SB 431542. After growth for 7 days, the cells were collected, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 . The cells were then stained with PE Mouse anti-Human Pax-6 and PerCP-Cy5.5 Mouse anti-Human Sox1 Cat. No. 561549 . The plot was derived from gated events based on light scattering characteristics for the neural induction. Flow cytometry was performed on a BD LSR II flow cytometry system. Show More Intracellular staining of Pax-6 in neural induction of human embryonic stem ES cells. H9 human ES cells WiCell, Madison, WI were cultured in mTeSR Stem Cell Technologies on plates coated with BD Matrigel hESC-qualified Matrix Cat. No. 354277 . Embryoid bodies EB were made and cultured in medium containing Knockout Serum Replacement Life Technologies without bFGF for 24 hours and then in medium containing 250 ng/ml human recombinant noggin R&D Systems and 10 mu M SB 431542 Tocris for 4 more days. The EB were then plated on BD Matrigel-coated plates and grown in medium with ITS supplement Sigma-Aldrich , noggin, and SB 431542. After growth for 7 days, the cells were collected, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 . The cells were then stained with PE Mouse anti-Human Pax-6 and PerCP-Cy5.5 Mouse anti-Human Sox1 Cat. No. 561549 . The plot was derived from gated events based on light scattering characteristics for the neural induction. Flow cytometry was performed on a BD LSR II flow cytometry system.