E.coli (ATCC 25922, 5 × 108 CFU) in logarithmic growth was fixed by 2.6% polyformaldehyde, 0.012% glutaraldehyde, and 30 mM phosphate buffer pH 7.4 for 15 min at room temperature and then on the rice for 30 min. The bacteria were washed with cold PBS three times and blocked with PBS solution containing 2% BSA+1% gelatin at room temperature for 30 minutes. The bacteria were given corresponding reagents for 1 h at room temperature. Next, cells were incubated with FITC-labeled Goat Anti-Rabbit IgG (1:200 dilution, Jackson ImmunoResearch Inc, West Grove, PA, USA) for 30 min. Finally, OD600 and fluorescence intensity were detected by Microplate Reader. The data was presented as fluorescence intensity of FITC-labeled rabbit anti-mouse IgG-Fc antibody/bacterial density (OD600).
Fitc labeled goat anti rabbit igg
Manufactured by Jackson ImmunoResearch
Sourced in United States
FITC-labeled goat anti-rabbit IgG is a secondary antibody used for detection in immunoassays. It binds to rabbit primary antibodies and is conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate) for visualization.
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Lab products found in correlation
Dmirb inverted microscope, by Leica camera (1 mentions)
Cy3 labeled anti mouse, by Jackson ImmunoResearch (1 mentions)
Fish skin gelatin, by Merck Group (1 mentions)
Epifluorescence microscope, by Zeiss (1 mentions)
Axiocam mrm ccd camera, by Zeiss (1 mentions)
Live dead assay kit, by Thermo Fisher Scientific (1 mentions)
Axiovision software, by Zeiss (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Anti nucleolin antibody, by Abcam (1 mentions)
A1 confocal microscope, by Nikon (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
9 protocols using fitc labeled goat anti rabbit igg
1
Quantify Antibody Binding to E. coli
2
Immunohistochemical Analysis of Colon Tissue
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Paraffin-embedded human colonoid and colon tissue sections were deparaffinized followed by antigen retrieval by boiling in citrate acid buffer (#S23045–500, Research products International Corporation) in a pressure cooker at 105°C Sections were blocked using 10% goat serum for 1 h at room temperature. Sections were stained with ZO-1, H3K4me1, and MPO by incubation with a rabbit anti-mouse ZO-1 (#61–7300, Invitrogen Life Technologies) antibody, a rabbit polyclonal anti-H3K4me1 antibody (#5326, Cell Signaling), and anti-MPO (#MPO-121-FITC, FabGennix International Incorporated) overnight at 4°C, followed by a FITC-labeled goat anti-rabbit IgG (#111-165-003, Jackson ImmunoResearch) antibody or Cy3-labeled anti-mouse (#115-165-003, Jackson ImmunoResearch) antibody at room temperature for 2 h. Sections were then mounted using Mounting Medium containing DAPI for nuclear counter-staining. Slides were scanned using the Apiro Versa 200 platform or observed using Leica DM IRB inverted microscope and images were recorded using an Echo/Revolve (CD230RevA) camera.
3
Immunofluorescence Antibody Labeling
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The following antibodies were purchased: FITC-conjugated F(ab′)2 fragment of donkey anti-mouse IgG (H + L) (Jackson ImmunoResearch, West Grove, PA), anti-CD5 (Q-20, goat polyclonal IgG for mouse CD5 at the N-terminus, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-NEU1 middle region antibody (rabbit polyclonal antibody, Aviva Systems Biology (San Diego, CA)), Anti-AIRE (rabbit polyclonal IgG against AIRE (KLH-conjugated synthetic peptide derived from human AIRE, purified by protein A (cross-reactive species: human, mouse, rat), from Bioss (Boston, MA)). The secondary antibody was FITC-labeled goat anti-rabbit IgG (Jackson ImmunoResearch) or Rhodamine (TRITC)-conjugated donkey anti-rabbit IgG (H + L) (Jackson ImmunoResearch). Rhodamine RedTM-X (R.R.)-anti-mouse IgG (R.R.-conjugated AffiniPure F(ab′)2 fragment of donkey anti-mouse IgG(H + L)) and R.R.-anti-mouse IgM (R.R.-conjugated AffiniPure F(ab′)2 fragment of donkey anti-mouse IgM, μ chain specific) were purchased from Jackson ImmunoResearch. For FITC-anti-CD5, the anti-CD5 antibody described above was conjugated to FITC, as previous described48 . An anti-mouse MHC class II (anti I-Ak, 10-2-16 (monoclonal antibody) cell line was obtained from the American Type Culture Collection (Manassas, VA), and the culture supernatant was purified, as described previously39 (link) and labeled with FITC, as described previously48 .
4
Immunostaining of Cultured Neurons
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For immunostaining, cultured neurons were fixed at indicated times in PBS containing 4% paraformaldehyde/4% sucrose for 20 minutes at 37°C. The neurons were then permeabilized with 0.1% Triton X-100 (Sigma)/PBS for 5 minutes and blocked with 0.5% fish skin gelatin (Sigma)/PBS for 1 hour, both at 27°C. The cells were then stained for somatodendritic markers using polyclonal rabbit anti-microtubule-associated protein 2 antibody (1:150; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C overnight, followed by incubation with fluorescein isothiocyanate (FITC)-labeled goat anti-rabbit IgG (1:200; Jackson Immuno Research Labs, Inc., West Grove, PA, USA) at 37°C for 2 hours. The samples were washed with PBS, dewaxed, and mounted on slide glasses. The stained samples were imaged under an epifluorescence microscope (Zeiss, Oberkochen, Germany) equipped with a 20 × objective lens. The images were obtained with Axiocam MRm CCD camera (Zeiss) and AxioVision software (Zeiss). The toxic effects of aspartic acid on the neurons were evaluated by Live/Dead Assay kit (L-3224; Molecular Probe, Eugene, OR, USA) and the percent of damaged neuron number/total neuron numbers was calculated[63 (link)]. Neuronal viability was evaluated and compared with cultured neurons not exposed to aspartic acid.
5
Immunofluorescence Staining of CAT-1, CAT-2, and iNOS
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The sections used for immunofluorescence staining were obtained from the same paraffin blocks and were deparaffinized with xylene solution. After rehydration with a graded series of ethanol in phosphate-buffered saline, 0.3% hydrogen peroxidase was added to block the intrinsic peroxidase activity, and after rinsing with 5% normal rat serum, the specimens were incubated overnight at 4°C with monoclonal antibodies against CAT-1, CAT-2, or iNOS (Santa Cruz Biotechnology, Santa Cruz, CA). FITC-labeled goat anti-rabbit IgG (1 : 200 dilution; Jackson ImmunoResearch, West Grove, PA) was used as the secondary antibody. The specimens were examined using fluorescence microscopy, and the same exposure was used to photograph all of the samples. All of the slices for the immunofluorescence staining of CAT-1, CAT-2, and iNOS were also counterstained with 4′,6-diamidino-2-phenylindole (DAPI).
6
Immunofluorescent Analysis of NF-κB
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Peritoneal macrophages were cultured in a 24-well plate and given corresponding reagents for the indicated time. Cells were then fixed with 4% PFA for 30 min at 37°C. After that, cells were permeabilized with PBS-T (0.1% Triton X-100 dissolved in PBS) for 10 min and blocked with 4% BSA in PBS. Next, cells were incubated with NF-κB antibody (1:100 dilution) overnight at 4°C, followed by staining with FITC-labeled Goat Anti-Rabbit IgG (1:200 dilution, Jackson ImmunoResearch Inc, West Grove, PA, USA) for 2 h. Finally, cells were stained for DAPI in dark for 5 min and the fluorescence was observed by microscope.
7
Tumor Targeting with DOX@UiO-66/Py-PGA-PEG-F3
8
Quantifying PEDV Viral Titers via IFA
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IFA was performed on Vero or M145 96-well monolayers. Infected wells were fixed in cold ethyl alcohol and polyclonal rabbit anti-PEDV nucleoprotein (NP) antiserum (South Dakota State University Animal Disease Research and Diagnostic Laboratory (SDSU)) was added at 1:500. Cells were rinsed and then incubated with fluorescein isothiocyanate (FITC) labeled goat anti-rabbit IgG (Jackson Immunoresearch) at a dilution of 1:50, and then read using a fluorescent microscope. Tissue culture infective dose (TCID50/mL) was calculated using the Spearman-Karber method.
9
Visualizing YAP and TAZ Nuclear Translocation
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To assess YAP's and TAZ's translocation from the cytoplasm into the nucleus, the MC3T3-E1 cells were incubated on the Polished Ti and SLA Ti disks for 24 h. The cells were fixed with 4% paraformaldehyde for 15 min and permeabilized with 0.5% Triton X-100 for 20 min. The nonspecific antibody interaction was blocked with goat serum for 30 min. The disks were incubated with rabbit monoclonal antibody to YAP (1:100 dilution; Cell Signaling Technology, Boston, MA, USA) and rabbit polyclonal antibody to TAZ (1:100 dilution; Abcam, Cambridge, UK) overnight at 4°C. Then they were incubated with a FITC-labeled goat anti-rabbit IgG (1:50 dilution; Jackson Immuno Research, Baltimore, MD, USA) to visualize the primary antibodies at 37°C for 50 min. The nuclei were counterstained with 4',6 diamidino-2-phenylindole (DAPI, Invitrogen) for 5 min. The disks were rinsed with PBS buffer in the interval of each of the two reagents. YAP's and TAZ's translocation was visualized and photographed (LSM 780 Spectral Confocal Laser Scanning microscope, Zeiss, Jena, Germany).
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