rabbit anti-gitrl antibody  (GeneTex)

Journal: The Journal of investigative dermatology

Article Title: Identification of Glucocorticoid-Induced TNF Receptor-Related Protein Ligand on Keratinocytes: Ligation by GITR Induces Keratinocyte Chemokine Production and Augments T-Cell Proliferation

doi: 10.1038/jid.2009.163

Figure Lengend Snippet: Skin punch biopsies obtained from (a) male Balb/C mice and (b) male C57Bl/6 mice were immunostained with anti-GITRL Ab (right panel, green) or with control IgG (left panel). The sections were counterstained with DAPI (blue). Representative data out of three independent experiments is shown (bar = 10 μm).

Article Snippet: Rabbit anti-mouse GITRL Ab, goat anti-mouse TARC Ab (clone N-20), and NF-κB p50 (clone D-17) Abs were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Control

Journal: The Journal of investigative dermatology

Article Title: Identification of Glucocorticoid-Induced TNF Receptor-Related Protein Ligand on Keratinocytes: Ligation by GITR Induces Keratinocyte Chemokine Production and Augments T-Cell Proliferation

doi: 10.1038/jid.2009.163

Figure Lengend Snippet: Keratinocytes regulate the expression of TARC (a and b), CTACK (c and d), MCP-1 (e and f), and MBD3 (g and h) mRNA levels after reverse stimulation through GITRL. Skin biopsies from male Balb/C mice were incubated for 6 hours (a, c, e, and g) or 24 hours (b, d, f, and h) in the presence or absence of GITR: Fc FP (1 or 10 μg ml−1) or Control: Fc FP, then harvested and analyzed for mRNA expression by RT-PCR using primers specific for TARC, CTACK, MCP-1, MBD3, and 18S RNA. Values indicate the mean±SEM of six independent experiments.

Article Snippet: Rabbit anti-mouse GITRL Ab, goat anti-mouse TARC Ab (clone N-20), and NF-κB p50 (clone D-17) Abs were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Expressing, Incubation, Control, Reverse Transcription Polymerase Chain Reaction

Journal: The Journal of investigative dermatology

Article Title: Identification of Glucocorticoid-Induced TNF Receptor-Related Protein Ligand on Keratinocytes: Ligation by GITR Induces Keratinocyte Chemokine Production and Augments T-Cell Proliferation

doi: 10.1038/jid.2009.163

Figure Lengend Snippet: (a) Cells were stained using PE-conjugated anti-GITRL Ab before and after fixing with 1% paraformaldehyde after overnight culture and analyzed by flow cytometry. The histogram represents one of five independent experiments. The black histogram denotes anti-GITRL Ab and the blue histogram denotes the isotype. (b) Cells were stained with PE-conjugated anti-GITRL Ab (red) and DAPI (blue) and analyzed by immunofluorescence (bar = 20 μm). (c) PAM 212 cells were incubated with GITR: Fc FP (1 or 10 μg ml−1) or with Control: Fc FP (10 μg ml−1) for 24 hours. Supernatants were harvested and TARC production was quantified by ELISA. Values indicate the mean±SEM of six independent experiments.

Article Snippet: Rabbit anti-mouse GITRL Ab, goat anti-mouse TARC Ab (clone N-20), and NF-κB p50 (clone D-17) Abs were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Staining, Flow Cytometry, Immunofluorescence, Incubation, Control, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Identification of Glucocorticoid-Induced TNF Receptor-Related Protein Ligand on Keratinocytes: Ligation by GITR Induces Keratinocyte Chemokine Production and Augments T-Cell Proliferation

doi: 10.1038/jid.2009.163

Figure Lengend Snippet: (a) Cells were stained using both directly and indirectly FITC-conjugated anti-GITRL Ab and analyzed by flow cytometry. The histogram represents one of five independent experiments. The black histogram denotes anti-GITRL Ab and the blue histogram denotes the isotype. (b) Cells were stained with FITC-conjugated anti-GITRL Ab (green) and DAPI (blue) and analyzed by immunofluorescence (bar = 20 μm). HEKs regulate the expression of (c) CTACK (d) and IL-8 mRNA level after reverse stimulation through GITRL. Human keratinocytes were incubated for 6 hours in the presence or absence of GITR: Fc FP (1 or 10 μg ml−1) or Control: Fc FP, then harvested and analyzed for mRNA expression by RT-PCR using primers specific for CTACK, IL-8, and 18S RNA. Values indicate the mean±the SEM of three independent experiments.

Article Snippet: Rabbit anti-mouse GITRL Ab, goat anti-mouse TARC Ab (clone N-20), and NF-κB p50 (clone D-17) Abs were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Staining, Flow Cytometry, Immunofluorescence, Expressing, Incubation, Control, Reverse Transcription Polymerase Chain Reaction

Journal: The Journal of investigative dermatology

Article Title: Identification of Glucocorticoid-Induced TNF Receptor-Related Protein Ligand on Keratinocytes: Ligation by GITR Induces Keratinocyte Chemokine Production and Augments T-Cell Proliferation

doi: 10.1038/jid.2009.163

Figure Lengend Snippet: Murine CD4+ T cells were isolated from mouse spleens and stimulated with anti-CD3 Ab in the presence or absence of keratinocytes. (a) Keratinocytes (1 × 104 well) enhance T-cell proliferation at increasing concentrations of anti-CD3 Ab (0.01–0.5 μg ml−1). (b) Keratinocytes were cultured with anti-GITRL blocking Ab (1 μg ml−1) or with mouse IgG1 isotype control for 1 hour before their use in proliferation assays.

Article Snippet: Rabbit anti-mouse GITRL Ab, goat anti-mouse TARC Ab (clone N-20), and NF-κB p50 (clone D-17) Abs were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Isolation, Cell Culture, Blocking Assay, Control