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Fitc and alexa 568 conjugated secondary antibodies

Manufactured by Thermo Fisher Scientific
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FITC-and Alexa 568-conjugated secondary antibodies are fluorescently labeled antibodies used in various immunodetection applications. These antibodies can bind to primary antibodies and emit fluorescent signals, enabling the visualization and detection of target proteins or other biomolecules in samples.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions) Sc 53253, by Santa Cruz Biotechnology (2 mentions) Eclipse e600 microscope, by Zeiss (2 mentions) Labtek 2 chambers, by Thermo Fisher Scientific (1 mentions) X81 fluorescence microscope, by Olympus (1 mentions)

Close spelling variants of this product

Alexa-568 secondary antibodies Alexa secondary antibodies FITC-conjugated antibodies Alexa 568

3 protocols using fitc and alexa 568 conjugated secondary antibodies

1

Immunofluorescence Staining of Epithelial Markers

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Cells were grown on coverslips coated with poly-lysine until 75% confluent. The cells were then washed twice in ice-cold phosphate buffered saline (PBS), fixed with 4% paraformaldehyde for 20 min at room temperature, permeabilized with 0.5% Triton in 1% glycine and then blocked using 0.5% BSA and 5% goat serum for 30 min at room temperature. Samples were then incubated with antibodies against P63 (Thermo Scientific, catalog # 703809, 1:200 dilution), and cytokeratin 14 (Santa Cruz Biotechnology, catalog #sc-53253, 1:200 dilution) in blocking solution overnight at 4 °C. Samples were then washed three times with PBS containing Tween 20 (PBST), followed by incubation with FITC-and Alexa 568- conjugated secondary antibodies (Invitrogen, 1:1000 dilution). Nuclei were also stained with 4', 6-diamidino-2-phenylindole (DAPI) (Invitrogen, 1:5000 dilution) before cells were mounted. Cells were observed using a Nikon Eclipse E600 microscope and a Zeiss confocal laser-scanning microscope.
Woappi Y., Altomare D., Creek K.E, & Pirisi L. (2020). Self-assembling 3D spheroid cultures of human neonatal keratinocytes have enhanced regenerative properties. Stem cell research, 49, 102048.
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2

Immunofluorescence Staining of Epithelial Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were grown on coverslips coated with poly-lysine until 75% confluent. The cells were then washed twice in ice-cold phosphate buffered saline (PBS), fixed with 4% paraformaldehyde for 20 min at room temperature, permeabilized with 0.5% Triton in 1% glycine and then blocked using 0.5% BSA and 5% goat serum for 30 min at room temperature. Samples were then incubated with antibodies against P63 (Thermo Scientific, catalog # 703809, 1:200 dilution), and cytokeratin 14 (Santa Cruz Biotechnology, catalog #sc-53253, 1:200 dilution) in blocking solution overnight at 4 °C. Samples were then washed three times with PBS containing Tween 20 (PBST), followed by incubation with FITC-and Alexa 568- conjugated secondary antibodies (Invitrogen, 1:1000 dilution). Nuclei were also stained with 4', 6-diamidino-2-phenylindole (DAPI) (Invitrogen, 1:5000 dilution) before cells were mounted. Cells were observed using a Nikon Eclipse E600 microscope and a Zeiss confocal laser-scanning microscope.
Woappi Y., Altomare D., Creek K.E, & Pirisi L. (2020). Self-assembling 3D spheroid cultures of human neonatal keratinocytes have enhanced regenerative properties. Stem cell research, 49, 102048.
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3

Immunofluorescence Analysis of Epithelial Markers

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HKc/HPV16-Ctrl and HKc/HPV16-Six1 were plated in Lab-Tek II chambers (Nalge Nunc International, Rochester, NY) for 24 h. Following treatment, fixation (with 2% paraformaldehyde in PBS, pH 7.2) and permeabilization (with 0.1% Triton X-100 in PBS), samples were incubated with antibodies against E-cadherin (at 1:200 dilution), fibronectin (at 1:200 dilution) and keratin 10 (at 1:100 dilution) overnight at 4°C. Samples were then washed three times with PBST, followed by incubation with FITC- and Alexa 568- conjugated secondary antibodies (at 1:1000 dilution, Invitrogen). Nuclei were stained with a 1:20,000 dilution of 4’, 6-diamidino-2-phenylindole (DAPI) (Invitrogen) before cells were mounted. Samples were observed using an Olympus X81 fluorescence microscope.
Xu H., Pirisi L, & Creek K.E. (2014). Six1 Overexpression at Early Stages of HPV16-mediated Transformation of Human Keratinocytes Promotes Differentiation Resistance and EMT. Virology, 474, 144-153.
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