Western Blotting (WB), Immunofluorescence (IF), BioImaging (BI), Intracellular Staining (ICS)
Marke
BD Pharmingen™
Produktmerkmale
Pax-6 is a member of the paired box (pax) gene family whose protein products are transcription factors involved in development. Pax family members share a highly conserved DNA binding domain that contains six alpha helices (paired domain)and a homeo box domain. Pax-6 has important roles in the development of the eye, nose, central nervous system, and pancreas. Defects in Pax-6 are responsible for various eye malformations including aniridia and Peters anomaly. The O18-1330 monoclonal antibody reacts with human Pax-6. Because the Pax-6 protein sequence is highly conserved among vertebrate species, cross-reactivity with other species is possible. LEFT: 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 mM 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 21 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 either Purified Mouse anti-Human Pax-6 antibody (solid line histrogram) or Purified Mouse IgG2a, k isotype control (dashed line, Cat. No. 554126). The second-step reagent was PE Goat Anti-Mouse Ig (Multiple Adsorption, Cat. No. 550589). Flow cytometry was performed on a BD LSR™ II flow cytometry system. RIGHT: Immunofluorescent staining of Pax-6 in human embryonic stem cell-derived rosettes. H9 human ES cells (WiCell, Madison, WI) passage 54 cultured in mTeSR™1 medium (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were differentiated towards a neural stem cell lineage. Cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1 % Triton™ X-100 [Perm Buffer III is also suitable], and stained with Purified Mouse anti-Human Pax-6 (pseudo-colored green) at 2.5 μg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies) and counter-staining was with Hoechst 33342 (Sigma-Aldrich, pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software. 561462 Rev. 2 Page 1 of 2
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Lagerung
4 °C
Informationen zur Lagerung
Store undiluted at 4°C.
Chambers, Fasano, Papapetrou, Tomishima, Sadelain, Studer: "Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling." in: Nature biotechnology, Vol. 27, Issue 3, pp. 275-80, (2009) (PubMed).
Osakada, Jin, Hirami, Ikeda, Danjyo, Watanabe, Sasai, Takahashi: "In vitro differentiation of retinal cells from human pluripotent stem cells by small-molecule induction." in: Journal of cell science, Vol. 122, Issue Pt 17, pp. 3169-79, (2009) (PubMed).
Cerf: "Transcription factors regulating beta-cell function." in: European journal of endocrinology / European Federation of Endocrine Societies, Vol. 155, Issue 5, pp. 671-9, (2006) (PubMed).
Glaser, Walton, Maas: "Genomic structure, evolutionary conservation and aniridia mutations in the human PAX6 gene." in: Nature genetics, Vol. 2, Issue 3, pp. 232-9, (1994) (PubMed).