Journal: Journal of Integrative Agriculture

Article Title: CDH1, a Novel Surface Marker of Spermatogonial Stem Cells in Sheep Testis

doi: 10.1016/s2095-3119(13)60689-9

Figure Lengend Snippet: Fig. 1 The map of the recombinant vector for expression of His-CDH1 fusion peptide. The CDH1 ectodomain was link to the His-tag in the pET-44a(+) vector. Expression of His-CDH1 fusion peptide was driven by T7 lac promoter.

Article Snippet: After blocking with 5% skim milk for 1 h at RT, the seminiferous tubules were incubated with CDH1 polyclonal antibody and anti-PLZF antibody (Santa Cruz) overnight at 4°C.

Techniques: Recombinant, Plasmid Preparation, Expressing

Journal: Journal of Integrative Agriculture

Article Title: CDH1, a Novel Surface Marker of Spermatogonial Stem Cells in Sheep Testis

doi: 10.1016/s2095-3119(13)60689-9

Figure Lengend Snippet: Fig. 2 Analysis of the optimal expressed condition of His-CDH1 fusion peptide by SDS-PAGE. M, protein molecular weight marker; 1, control; 2-9, the sample induced by IPTG at 1, 1.5, 2, 2.5, 3, 4, 5 and 6 h, respectively.

Article Snippet: After blocking with 5% skim milk for 1 h at RT, the seminiferous tubules were incubated with CDH1 polyclonal antibody and anti-PLZF antibody (Santa Cruz) overnight at 4°C.

Techniques: SDS Page, Molecular Weight, Marker, Control

Journal: Journal of Integrative Agriculture

Article Title: CDH1, a Novel Surface Marker of Spermatogonial Stem Cells in Sheep Testis

doi: 10.1016/s2095-3119(13)60689-9

Figure Lengend Snippet: Fig. 3 Western blot analysis of the condition for purifying His- CDH1 peptide using Ni-NTA chromatography. 1, prestained protein marker; 2, the recovery fluid with lysate supernatant; 3, the recovery fluid with 20 mmol imidazol; 4-5, the recovery fluid with 100 mmol imidazol; 6-7, the recovery fluid with 250 mmol imidazol; 8-9, the recovery fluid with 500 mmol imidazol; 10, the recovery fluid with 1 000 mmol imidazol.

Article Snippet: After blocking with 5% skim milk for 1 h at RT, the seminiferous tubules were incubated with CDH1 polyclonal antibody and anti-PLZF antibody (Santa Cruz) overnight at 4°C.

Techniques: Western Blot, Chromatography, Marker

Journal: Journal of Integrative Agriculture

Article Title: CDH1, a Novel Surface Marker of Spermatogonial Stem Cells in Sheep Testis

doi: 10.1016/s2095-3119(13)60689-9

Figure Lengend Snippet: Fig. 4 Ability of the sheep CDH1 polyclonal antibodies was detected by Western blotting. Total protein was from sheep testis (T), kinedy (K), pulmo (P) and liver (L). There were three bands in 135, 120 and 80 kDa.

Article Snippet: After blocking with 5% skim milk for 1 h at RT, the seminiferous tubules were incubated with CDH1 polyclonal antibody and anti-PLZF antibody (Santa Cruz) overnight at 4°C.

Techniques: Western Blot

Journal: Journal of Integrative Agriculture

Article Title: CDH1, a Novel Surface Marker of Spermatogonial Stem Cells in Sheep Testis

doi: 10.1016/s2095-3119(13)60689-9

Figure Lengend Snippet: Fig. 5 Distribution patterns of CDH1 and PLZF in cross sections of 5-mon-old sheep testis. A, immunohistochemistry examination of PLZF expressing cells within cross sections of seminiferous tubules from 5-mon-old sheep testis. B, immunohistochemistry examination of CDH1 in the same section. C, the nuclei of the cell sections were stained by Hoechst 33342. D, merged image of A, B and C. Some PLZF- and CDH1-positive cells comprising single (diamond arrows) and paired spermatogonia (arrows) were localized at the basement membrane of the seminiferous tubules. Scale bars are 40 μm.

Article Snippet: After blocking with 5% skim milk for 1 h at RT, the seminiferous tubules were incubated with CDH1 polyclonal antibody and anti-PLZF antibody (Santa Cruz) overnight at 4°C.

Techniques: Immunohistochemistry, Expressing, Staining, Membrane

Journal: Journal of Integrative Agriculture

Article Title: CDH1, a Novel Surface Marker of Spermatogonial Stem Cells in Sheep Testis

doi: 10.1016/s2095-3119(13)60689-9

Figure Lengend Snippet: Fig. 6 Whole-mount immunohistochemistry of 5-mon-old sheep seminiferous tubules. The CDH1-positive cells were small in number, there are single cells attached the basement membrane of the seminiferous tubules (arrows), and there are paired cells attached each other (diamond arrow). Scale bar is 200 μm.

Article Snippet: After blocking with 5% skim milk for 1 h at RT, the seminiferous tubules were incubated with CDH1 polyclonal antibody and anti-PLZF antibody (Santa Cruz) overnight at 4°C.

Techniques: Immunohistochemistry, Membrane

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: A virus-like particle-based anti-nerve growth factor vaccine reduces inflammatory hyperalgesia: potential long-term therapy for chronic pain.

doi: 10.4049/jimmunol.1000030

Figure Lengend Snippet: FIGURE 1. Production and functional analysis of the rNGFQb vaccine. A, Production of rNGF. Murine NGF was expressed in HEK293T cells. Antipentahistidine immunoblot analysis of HEK293T lysates (first lane) and cell-culture supernatants (second lane) revealed expression of the NGF proprotein by HEK293T cells and secretion of the processed 14.7-kDa mature form of NGF into the cell-culture supernatant. Coomassie staining of NGF purified from supernatants by affinity chromatography confirmed homogeneity (third lane). B, Recognition of NGF by anti-NGF Abs. Recombinantly expressed NGF (white circles) was compared with ex vivo- purified NGF (black squares). Different dilutions of both proteins were applied to ELISA plates coated with anti-NGF mAb. Bound NGF was detected with polyclonal anti-NGF Abs. Averages of triplicates 6 SEM. C, NGF receptor binding. The NGF-specific receptor TrkA was coated onto ELISA plates and descending concentrations of both proteins applied. Receptor bound NGF was detected with polyclonal NGF Abs as in B. Averages of triplicates 6 SEM. D, Bioactivity of NGF. The NGF re- sponsive cell line TF-1 was stimulated with descending concentrations of NGF and proliferation determined by BrdU incorporation during DNA synthesis. Averages of triplicates 6 SEM. E, Analysis of NGFQb vaccine. NGF was cross-linked to VLPs with a heterobifunctional cross-linker. An immunoblot of NGF (left lane) or the NGFQb conjugate vaccine (right lane) analyzed with an anti-pentahistidine–specific Ab showed a dominant band appearing at 29 kDa corresponding to a Qb monomer (14.2 kDa) conjugated to one molecule NGF (14.7 kDa). Higher molecular mass bands correspond to Qb-oligomers linked to one NGF molecule. F, Binding of TrkA to NGFQb. Ascending concentrations of Qb (filled squares) or NGFQb (open circles) were immobilized on an ELISA plate coated with anti-Qb mAb. Bound vaccine was incubated with TrkA re- ceptor containing a human Fc domain. Binding of receptor to NGF dis- played on VLP was detected with peroxidase-conjugated goat anti-human Abs. Averages of triplicates 6 SEM. LY, lysates; SN, supernatants.

Article Snippet: Then, plates were washed six times with PBS-T and incubated with a 1:500 dilution of a sheep anti-NGF polyclonal Ab (AbD Serotec) for 1 h at RT.

Techniques: Functional Assay, Western Blot, Cell Culture, Expressing, Staining, Chromatography, Ex Vivo, Enzyme-linked Immunosorbent Assay, Binding Assay, BrdU Incorporation Assay, DNA Synthesis, Incubation