goat anti rabbit horseradish peroxidase conjugate  (Thermo Fisher)

Journal: Clinical and Experimental Immunology

Article Title: Relevance of differential immunogenicity of human and mouse recombinant desmoglein-3 for the induction of Acantholytic autoantibodies in mice

doi: 10.1046/j.1365-2249.2003.02135.x

Figure Lengend Snippet: Reactivity of anti-hdsg3 Abs with cells expressing mdsg3. The 293 cells were transfected with pSR α neo vector containing full-length mdsg3 cDNA, and stable cell lines were established (293 mdsg3). Cells were analysed by flow cytometry for the expression of mdsg3 by staining with rabbit anti-hdsg3 antibody (1 : 100), followed by FITC- conjugated goat antirabbit IgG (1 : 100). Propidium iodide was used to distinguish live from dead cells. — , 293 cells; ···, 293 mdsg3 cells.

Article Snippet: Blots were treated for 1 h with a blocking buffer (5% non-fat dry milk in 200 m m NaCl with 0·1% Tween-20), and then incubated for 1 h each with the following reagents in succession with washing between each step: 1 : 2000 diluted rabbit anti-hdsg3 antibody [ ], and 1 : 3000 diluted HRP-conjugated goat antirabbit IgG (Caltag Laboratories, San Francisco, CA, USA).

Techniques: Expressing, Transfection, Plasmid Preparation, Stable Transfection, Flow Cytometry, Cytometry, Staining

Journal: Oncotarget

Article Title: C-terminus of MUC16 activates Wnt signaling pathway through its interaction with β-catenin to promote tumorigenesis and metastasis

doi: 10.18632/oncotarget.9191

Figure Lengend Snippet: MUC16/MUC16C interacts with β-catenin A. Expression profile of MUC16 in different cell lines. The cell lysates were analyzed by Western blot with the self-made mouse anti-MUC16 (40 μg input) or mouse anti-β-actin (8 μg input) antibody respectively. B. Ectopically expressed MUC16C interacts with endogenous β-catenin. HeLa cells were transfected with pcDNA3.3-HA-MUC16C or the empty vector as a control. At 24 h post-transfection, cells were lysed and subjected to immunoprecipitation with mouse anti-HA antibody, followed by Western blot with mouse anti-HA and rabbit anti-β-catenin antibodies separately. C. MUC16 and β-catenin interact with each other in vivo. Lysates of SKBR-3 cells were subjected to immunoprecipitation with the control IgG, rabbit anti-β-catenin and mouse anti-MUC16 antibodies respectively. The precipitates were then detected with indicated antibodies. D. C-terminus of β-catenin is essential for its interaction with MUC16. For the domain mapping experiment, structures of deletion mutants of β-catenin are shown on the top of the panel. Functional domains of β-catenin are indicated above the schema; the remaining fragments of each deletion mutant are shown in the diagram. HEK293T cells were co-transfected with pcDNA3.3-HA-MUC16C and different MYC-β-catenin mutants or the empty vector as a control. At 24 h post-transfection, cells were lysed and subjected to immunoprecipitation with rabbit anti-MYC antibody, followed by Western blot with mouse anti-HA or anti-MYC antibody.

Article Snippet: Mouse anti-β-actin antibody was purchased from Santa Cruz Biotechnology, Inc. Goat-anti-mouse and goat anti-rabbit immunoglobulin G (IgG) horseradish peroxidase conjugates were purchased from Pierce Biotechnology, Inc.

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation, In Vivo, Functional Assay, Mutagenesis

Journal: PLoS Neglected Tropical Diseases

Article Title: Pharmacokinetics of Naja sumatrana (Equatorial Spitting Cobra) Venom and Its Major Toxins in Experimentally Envenomed Rabbits

doi: 10.1371/journal.pntd.0002890

Figure Lengend Snippet: Immunological cross reactions between N. sumatrana venom toxins as analyzed by immunoblotting. Venom toxins (10 µg each of phospholipase A 2 , neurotoxin and cardiotoxin) was electrophoresed on a SDS-PAGE gel (15%, reducing condition), and electro-transferred to a PVDF membrane. This was followed by subsequent incubation with primary antibody (anti-PLA 2 IgG, anti-NTX IgG and anti-CTX IgG (dilution of 1: 500) and goat anti-rabbit IgG-HRP (dilution of 1∶1000). Substrate solution (Novex HRP Chromogenic Substrate (TMB), Invitrogen) was added for colorimetric development.

Article Snippet: Anti-PLA2 IgG, anti-NTX IgG or anti-CTX IgG (dilution of 1∶500 in TBS-Tween) was added to the PVDF membrane followed by incubation with Goat anti-rabbit IgG horseradish peroxidase conjugate (dilutions of 1∶1000) for 1 h. The chromogenic detection of the protein bands on the PVDF membrane was carried out by addition of the substrate solution (Novex HRP Chromogenic Substrate (TMB), Invitrogen).

Techniques: SDS Page, Incubation, Proximity Ligation Assay

Journal: Scientific Reports

Article Title: Antiviral Phosphorodiamidate Morpholino Oligomers are Protective against Chikungunya Virus Infection on Cell-based and Murine Models

doi: 10.1038/srep12727

Figure Lengend Snippet: Histological analysis of mice pre-treated with CPMO1v. BALB/c mice (6-day old, n = 6) were i.p injected with 15 μg/g CPMO1v or sCPMO1v. CHIKV infection and treatment regimen is carried out as aforementioned in Fig. 5 . All mice were sacrificed at day 7 p.i. and their hind limbs and liver were harvested, fixed with formalin, ethanol-dehydrated and paraffin embedded in sections for ( a–h ) H E staining or ( i–p ) IHC staining using primary rabbit anti-CHIKV E2 IgG followed by secondary goat anti-rabbit HRP IgG. ( d ) ( f ) Arrow indicates smudgy nuclei inclusions while arrowhead indicates multi-nucleated inclusions of pale necrotic hepatocytes. ( k , l ) Arrow indicates positive CHIKV antigen staining. Images were viewed and captured at 400× under Olympus microscope. Representative images at 50 μm scale are shown.

Article Snippet: For IHC, tissue samples were labeled with primary rabbit E2 antibody diluted to 1:100, followed by a secondary goat anti-rabbit HRP conjugate (Thermoscientific).

Techniques: Mouse Assay, Injection, Infection, Staining, Immunohistochemistry, Microscopy