For immunofluorescence experiments, HKs were cultured directly on glass slides (Ibidi) until confluent, as described above. Live cells were labeled with monoclonal antibodies at a concentration of 10 μg/ml for 30 min at 4°C. Cells were then fixed on ice using 3.7% paraformaldehyde for 10 min followed by permeabilization in 0.1% triton for 10 min at room temperature. Goat anti-human IgG cross-adsorbed secondary (Life Technologies) was then used prior to mounting with prolong gold containing Dapi (Life Technologies). Widefield images were acquired using a DMRXA2 microscope (Leica, Wetzler, Germany) equipped with a 63X/1.40 NA oil immersion objective. Images were acquired with an ORCA digital camera (Hamamatsu Photonics, Bridgewater, NJ). Analysis was preformed using Fiji ImageJ.
Goat anti human igg cross adsorbed secondary
Manufactured by Thermo Fisher Scientific
Goat anti-human IgG cross-adsorbed secondary is a secondary antibody used in various immunoassay techniques. It is generated by immunizing goats with human IgG and is then cross-adsorbed to remove cross-reactivity with other species. This secondary antibody can be used to detect and quantify human IgG in samples.
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2 protocols using goat anti human igg cross adsorbed secondary
1
Immunofluorescence Imaging of Cultured Cells
2
Immunofluorescence Imaging of Cultured Cells
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For immunofluorescence experiments, HKs were cultured directly on glass slides (Ibidi) until confluent, as described above. Live cells were labeled with monoclonal antibodies at a concentration of 10 μg/ml for 30 min at 4°C. Cells were then fixed on ice using 3.7% paraformaldehyde for 10 min followed by permeabilization in 0.1% triton for 10 min at room temperature. Goat anti-human IgG cross-adsorbed secondary (Life Technologies) was then used prior to mounting with prolong gold containing Dapi (Life Technologies). Widefield images were acquired using a DMRXA2 microscope (Leica, Wetzler, Germany) equipped with a 63X/1.40 NA oil immersion objective. Images were acquired with an ORCA digital camera (Hamamatsu Photonics, Bridgewater, NJ). Analysis was preformed using Fiji ImageJ.
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