igg h 270  (Millipore)

Journal: Frontiers in Cell and Developmental Biology

Article Title: The antigenic determinant that defines thymic nurse cells is expressed by thymic epithelial progenitor cells

doi: 10.3389/fcell.2014.00013

Figure Lengend Snippet: Thymic embryonic sections stained with pH91 and anti-Foxn1 antibody. (A) shows embryonic staining with anti-Foxn1 antibody (magenta), left panel; along with pH91 (green) right panel of E11.5 mouse thymic primordium. Expression of Foxn1 is localized toward the ventral region of the embryo whereas pH91 expression is seen throughout the entire thymic and para-thyroid primordium. The merged image (lower panel) and inset (enlarged) show pH91 + cells in association with Foxn1 + cells (arrows). The insets of the upper left (inset) and upper right (inset) panels show secondary antibody control for Foxn1 antibody (magenta) and IgG2a isotope control for pH91 (green). (B) shows staining of E12.5 thymic anlagen with anti-Foxn1 antibody, left panel (magenta) and pH91, right panel (green). The merged image (lower panel) and inset (enlarged) show an association of Foxn1 + with pH91 + cells (arrows). Magnification 40×. (C) shows the relative percentage of the various phenotypic subtypes present at E11.5 and E12.5 respectively. The pH91 − /Foxn1 + phenotype shows a significant increase ( * P < 0.001) between E11.5 and E12.5, ( n = 6). The pH91 + /Foxn1 + subset show statistical significance ( ** P < 0.05) between E11.5 and E12.5 ( n = 6). Five-hundred and fifty embryonic cells were counted for (C) . The images are representative of three independent expriments.

Article Snippet: Primary antibodies used were as follows: rat anti-mouse pH91 monoclonal antibody (IgG2a), K8 TROMA-I (IgG2a) (Developmental Studies Hybridoma Bank, Iowa City, IA), chicken anti-mouse K8 polyclonal antibody (IgY) (Abcam, Cambridge, MA), goat anti-rabbit K5 polyclonal antibody PRB-160B (IgG) (Covance, Princeton, NJ), rabbit anti-goat Δ Np63 (N-16): sc-8609 (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-Foxn1 polyclonal antibody (IgG) H-270 (Santa Cruz Biotechnology), antibody FITC-conjugated anti-mouse MHC class II (Miltenyi Biotech), biotinylated anti-mouse αβ TCR (BD Pharmingen, San Jose, CA), APC-conjugated CD4 (BD Pharmingen), PE-conjugated CD8 (BD Pharmingen), FITC-conjugated Thy 1.2 (BD Pharmingen), FITC-conjugated rat IgG2a isotype control (BD Pharmingen), and TRITC-conjugated rabbit IgG2a isotype control (BD Pharmingen).

Techniques: Staining, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: The antigenic determinant that defines thymic nurse cells is expressed by thymic epithelial progenitor cells

doi: 10.3389/fcell.2014.00013

Figure Lengend Snippet: Expression of Foxn1 and the pH91-antigen in embryonic sections . Thymic section at embryonic stage E13.5 (A) , E16.5 (B) , and E18.5 (C) were stained with anti-Foxn1 antibody (magenta) and pH91mAb (green). The merged images and insets ( A–C) , E13.5; E16.5; and E18.5 show an association of pH91 + cells with Foxn1 + cells. Magnification 20×. (D) shows thymic section co-stained with K5 (red) and pH91 (green), The asterisks highlight pH91 + cells within the thymic cortex. M indicates the location of the medulla. Magnification 40×. (Ea–c) shows the expression of Foxn1 and pH91 in thymic tissue section 2 weeks after birth. Sections were stained with anti-Foxn1 (magenta); pH91 (green). (Ec) show merged image and insets of pH91 + cells in association with Foxn1 (Arrow). Magnification 40×. (F) shows the expression of Foxn1 in pH91 + TNC ex vivo : Using antibody against Foxn1, expression was observed ex vivo in the TNCs. Foxn1 nuclear localization is seen only within TNC nucleus (Fd) . pH91 staining is detected (Fe) , DAPI nuclear staining (Ff) and the merged image is represented in (Fg) . Thymocytes do not show Foxn1 expression. (G) shows secondary antibody controls for Foxn1 (Gh) and pH91 (Gi) , All images represent 3 independent experiments. Magnification 40×.

Article Snippet: Primary antibodies used were as follows: rat anti-mouse pH91 monoclonal antibody (IgG2a), K8 TROMA-I (IgG2a) (Developmental Studies Hybridoma Bank, Iowa City, IA), chicken anti-mouse K8 polyclonal antibody (IgY) (Abcam, Cambridge, MA), goat anti-rabbit K5 polyclonal antibody PRB-160B (IgG) (Covance, Princeton, NJ), rabbit anti-goat Δ Np63 (N-16): sc-8609 (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-Foxn1 polyclonal antibody (IgG) H-270 (Santa Cruz Biotechnology), antibody FITC-conjugated anti-mouse MHC class II (Miltenyi Biotech), biotinylated anti-mouse αβ TCR (BD Pharmingen, San Jose, CA), APC-conjugated CD4 (BD Pharmingen), PE-conjugated CD8 (BD Pharmingen), FITC-conjugated Thy 1.2 (BD Pharmingen), FITC-conjugated rat IgG2a isotype control (BD Pharmingen), and TRITC-conjugated rabbit IgG2a isotype control (BD Pharmingen).

Techniques: Expressing, Staining, Ex Vivo

Journal: Frontiers in Cell and Developmental Biology

Article Title: The antigenic determinant that defines thymic nurse cells is expressed by thymic epithelial progenitor cells

doi: 10.3389/fcell.2014.00013

Figure Lengend Snippet: Expression of Foxn1 and p63 in freshly isolated pH91 + multicellular TNCs ex vivo . (Aa) shows ex vivo TNC stained with anti-Foxn1 antibody. (Ab) shows ex vivo TNC stained with anti-p63 antibody. (Ac) shows ex vivo TNC stained with pH91 mAb antibody. (Ad) DAPI stained nuclei. (Ae) shows merged image of all three stains. (Bf–j) represents antibody controls for Foxn1, p63, and pH91. Magnifications 40×.

Article Snippet: Primary antibodies used were as follows: rat anti-mouse pH91 monoclonal antibody (IgG2a), K8 TROMA-I (IgG2a) (Developmental Studies Hybridoma Bank, Iowa City, IA), chicken anti-mouse K8 polyclonal antibody (IgY) (Abcam, Cambridge, MA), goat anti-rabbit K5 polyclonal antibody PRB-160B (IgG) (Covance, Princeton, NJ), rabbit anti-goat Δ Np63 (N-16): sc-8609 (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-Foxn1 polyclonal antibody (IgG) H-270 (Santa Cruz Biotechnology), antibody FITC-conjugated anti-mouse MHC class II (Miltenyi Biotech), biotinylated anti-mouse αβ TCR (BD Pharmingen, San Jose, CA), APC-conjugated CD4 (BD Pharmingen), PE-conjugated CD8 (BD Pharmingen), FITC-conjugated Thy 1.2 (BD Pharmingen), FITC-conjugated rat IgG2a isotype control (BD Pharmingen), and TRITC-conjugated rabbit IgG2a isotype control (BD Pharmingen).

Techniques: Expressing, Isolation, Ex Vivo, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: The antigenic determinant that defines thymic nurse cells is expressed by thymic epithelial progenitor cells

doi: 10.3389/fcell.2014.00013

Figure Lengend Snippet: Quantification of pH91 + multicellular TNCs expressing the transcription factors Foxn1 and p63 . pH91 + multicellular TNCs were isolated from, thymi from C57BL/6 mice at embryonic stages of development E13.5, E16.5, and E18.5, cyto-spun onto slide and quantified manually with respect to expression of Foxn1, p63 and pH91. (A) shows the embryonic expression of p63 and Foxn1 by pH91 + cells. (B) shows expression of p53 and Foxn1 by freshly isolated TNCs ex vivo . These data represent the results (mean ± standard deviation) collected from three independent experiments with three animals per group ( n = 9). * P < 0.05 compared E13.5 with E16.5. ** P < 0.01 compared E13.5 and E18.5. Statistical significance ( t -test).

Article Snippet: Primary antibodies used were as follows: rat anti-mouse pH91 monoclonal antibody (IgG2a), K8 TROMA-I (IgG2a) (Developmental Studies Hybridoma Bank, Iowa City, IA), chicken anti-mouse K8 polyclonal antibody (IgY) (Abcam, Cambridge, MA), goat anti-rabbit K5 polyclonal antibody PRB-160B (IgG) (Covance, Princeton, NJ), rabbit anti-goat Δ Np63 (N-16): sc-8609 (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-Foxn1 polyclonal antibody (IgG) H-270 (Santa Cruz Biotechnology), antibody FITC-conjugated anti-mouse MHC class II (Miltenyi Biotech), biotinylated anti-mouse αβ TCR (BD Pharmingen, San Jose, CA), APC-conjugated CD4 (BD Pharmingen), PE-conjugated CD8 (BD Pharmingen), FITC-conjugated Thy 1.2 (BD Pharmingen), FITC-conjugated rat IgG2a isotype control (BD Pharmingen), and TRITC-conjugated rabbit IgG2a isotype control (BD Pharmingen).

Techniques: Expressing, Isolation, Ex Vivo, Standard Deviation