Ovarian sections of immunized BALB/c and unimmunized wild mice were obtained from BIOSERV Analytik (Rostock, Germany). Ovarian sections were deparaffinized and subjected to antigen retrieval using sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0). Sections were blocked with 10% goat serum blocking solution (Life Technologies, Frederick, Maryland, USA) at RT for 1 h, washed with PBS, and incubated with 1X mouse-on-mouse IgG blocking solution (Invitrogen, Thermo Fisher Scientific) at RT for 1 h. Sections of unimmunized wild mice were incubated with a 1:20 dilution of serum samples from vaccinated BALB/c mice and kept at 4°C overnight. Sections of immunized BALB/c mice were incubated with a blocking solution (without the addition of any serum samples), washed, and incubated with 10 µg/mL fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG secondary antibody (Invitrogen, Thermo Fisher Scientific) at 37°C for 1.5 h. After washing, slides were mounted with the DABCO mounting medium (25 mg/mL DABCO, 90% glycerol, and 10% PBS; pH 8.5) and observed under a fluorescence microscope.
Fitc conjugated goat anti mouse igg secondary antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The (FITC)-conjugated goat anti-mouse IgG secondary antibody is a laboratory reagent used to detect and visualize mouse immunoglobulin G (IgG) in various immunoassay applications. The antibody is conjugated with the fluorescent dye Fluorescein isothiocyanate (FITC), allowing for the detection and quantification of target proteins through fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Goat serum blocking solution, by Thermo Fisher Scientific (1 mentions)
Mouse on mouse igg blocking solution, by Thermo Fisher Scientific (1 mentions)
Dapi containing solution, by Vector Laboratories (1 mentions)
Mouse anti nf κb p65, by Cell Signaling Technology (1 mentions)
Infinite f200 pro, by Tecan (1 mentions)
Normal goat serum (ngs), by Nichirei Biosciences (1 mentions)
Ab5417, by Abcam (1 mentions)
Triton x 100, by Thermo Fisher Scientific (1 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions)
Confocal microscope, by Leica (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Rabbit anti cd31 antibody, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Eclipse te2000 u, by Nikon (1 mentions)
Mouse anti cd34 antibody, by Abcam (1 mentions)
6 protocols using fitc conjugated goat anti mouse igg secondary antibody
1
Ovarian Immunohistochemistry in Mice
2
NF-κB p65 Translocation Assay in BV2 Cells
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To detect NF-κB p65 translocation, BV2 microglial cells were rinsed twice with PBS and fixed with 4% paraformaldehyde solution for 10 min, permeabilized with 0.1% (v/v) Triton-X100 for 20 min and blocked with 2% (w/v) BSA for 1 h. Cells were then sequentially incubated with mouse anti-NF-κB p65 (Cell Signaling Technology) overnight at 4 °C, and FITC-conjugated goat anti-mouse IgG secondary antibody (Invitrogen) for 45 min. The cells were then mounted in a DAPI-containing solution (Vector Laboratories, CA, USA), and images were captured by fluorescence microscopy (BX-51, Olympus Corp.).
3
Quantitative Analysis of Laminin Adsorption
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Recombinant Human laminin-332 (Ln; ReproCELL, Yokohama, Japan) was used as the model protein. All procedures for the adsorption assay were performed at room temperature (RT). Ti64 plates (64 and 64HT) were immersed in 0.5 µg/mL diluted Ln in phosphate-buffered saline (PBS) for 1 h. After rinsing off the unbound protein with PBS, the plates were treated with 10% normal goat serum (Nichirei Bioscience, Tokyo, Japan) in PBS for 10 min to prevent nonspecific adsorption of antibodies. The plates were then incubated with 4 µg/mL diluted mouse anti-human Ln monoclonal antibody (P3H9-2, Santa Cruz Biotechnology, Dallas, TX, USA) in PBS for 1 h. After rinsing off the unbound antibodies with PBS, the plates were treated with 10 µg/mL diluted fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG secondary antibody (Invitrogen, Carlsbad, CA, USA) in PBS for 30 min. The plates were then rinsed with PBS, and the fluorescent intensity was measured at an excitation wavelength of 485 nm and an emission wavelength of 535 nm using a microplate reader (Infinite F200PRO; Tecan, Salzburg, Austria) (n = 6 for each group).
4
Immunohistochemical Analysis of Rhodopsin
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Sections of samples embedded in paraffin were deparaffinized in a xylene ethanol series, placed in Tris-EDTA buffer for antigen retrieval (10 mM Tris, 1 mM EDTA, 0.05% Tween, pH = 9.0), and then blocked in 5% bovine serum albumin. Sections were immunostained for rhodopsin (rod photoreceptors) (Ab 5417; Abcam, United Kingdom; diluted 1:100). Detection of the primary antibodies was performed using fluorescein isothiocyanate (FITC)–conjugated goat anti-mouse IgG secondary antibody (F-2761; Thermo Fisher Scientific, United States; diluted 1:400). Nuclei were detected using 4′,6′-diamino-2-phenyl inodole (DAPI), which was included in the mounting solution (Solarbio; Beijing, China).
5
Immunofluorescence Assay for PTEN in Lung Cancer
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ED1 and HOP62 lung cancer cells with stable knockdown of USP18 were cultured on coverslips (Fisherbrand). After 24 hours, cells were washed (three times for five minutes each with PBS) and fixed with 4% paraformaldehyde (PFA) (Electron Microscopy Sciences) for 20 minutes at room temperature. Cells were washed again and permeabilized with 0.1% TritonX-100 (Fisherbrand) for 5 minutes at room temperature before blocking with 3% bovine serum albumin (PBS-BSA) for 1 hour at room temperature. Cells were incubated with a murine monoclonal antibody that recognized PTEN (Thermo Scientific #32-5800) or a murine monoclonal antibody isotype control (Biolegend #401602) diluted at 1:10 in 3% PBS-BSA and rabbit monoclonal anti-Sodium Potassium ATPase (Alexa Fluor 647) antibody (Abcam #198367) diluted at 1:100 in 3% PBS-BSA overnight at 4°C. The following day, cells were washed and incubated with a FITC-conjugated goat anti-mouse IgG secondary antibody (Thermo Scientific #F2761) diluted at 1:1000 in 3% PBS-BSA for 1 hour in the dark at room temperature. Cells were washed before mounting on coverslips (Fisherbrand) with Prolong Gold Anti-Fade Reagent with DAPI (Thermo Scientific). Fluorescence was examined using a confocal microscope (Leica Microsystems) and quantitated by using ImageJ (Fiji) software (version 2.0.0-rc-43/1.50e).
6
Immunofluorescence Analysis of Rat Skin Tissue
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The rat skin tissue samples collected from the wound area were mounted in OCT embedding compound and frozen at −80°C. The tissue block was cut into horizontal and cross sections with 5 µm thickness by a cryostat. The tissue sections were incubated either with mouse anti-CD34 antibody (1:350, Abcam, Cambridge, MA) and rabbit anti-CD31 antibody (1:350, Abcam, Cambridge, MA), or mouse anti-Engrailed-1antibody (1:350, Abcam, Cambridge, MA) and rabbit anti-neurofibromin antibody (1:350, Abcam, Cambridge, MA) overnight at 4°C. In the next morning, the sections were washed three times in PBS for 5 min before being incubated for 2 h at room temperature with FITC-conjugated goat anti-mouse IgG secondary antibody (1:500, ThermoFisher Scientific, Waltham, MA) for CD34 and engrailied-1 testing, and Cy3-conjugated goat anti-rabbit IgG second antibody (1:500, Millipore Sigma, Burlington, MA) for CD31 and neurofibromin testing. Slides were washed with PBS for three times, counterstained with 4′,6-diamidino-2-phenylindole (DAPI; 1 µg/ml, Sigma, St. Louis, MO) nuclear stain, and photographed by a fluorescence microscope (Nikon eclipse, TE2000-U).
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