Cells were cultured on microscope coverslips under hypoxic or normoxic conditions. After 16 h, slides were washed with phosphate-buffered saline (PBS), fixed for 20 min in 4% formaldehyde, and re-hydrated in PBS. After blocking for 45 min in 5% bovine serum albumin in PBS, the slides were incubated overnight at 4℃ with polyclonal rabbit anti-HIF-1α, -E-cadherin, and -vimentin antibodies (1:100; Cell Signaling Technology), washed with PBS, and incubated for 45 min with Alexa 488-labeled goat anti-rabbit antibodies or Alexa 568-labeled donkey anti-mouse antibodies (1:250; Molecular Probes, Inc., Eugene, OR, USA). The slides were washed with PBS and mounted using Duolink mounting medium with DAPI (Olink Bioscience, Sweden) and then analyzed using a LSM700 confocal microscope (Carl Zeiss Vision, Germany).
Alexa 488 labeled goat anti rabbit antibodies
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa 488-labeled goat anti-rabbit antibodies are a secondary antibody conjugate used for the detection and visualization of primary rabbit antibodies in various immunoassays and imaging techniques. The antibodies are labeled with the fluorescent dye Alexa Fluor 488, which emits green fluorescence upon excitation.
Automatically generated - may contain errors
2 protocols using alexa 488 labeled goat anti rabbit antibodies
1
Hypoxia-induced HIF-1α and EMT markers
2
Non-Thermal Plasma Regulation of Cell Adhesion
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Cells were cultured on microscope coverslips (Thermo Fisher Scientific, Rochester, NY, USA) and treated with gas (He+O2) only, 2, or 4 kV of NTP, respectively. After 24 h, the slides were washed with PBS, fixed for 20 min in 3.7% formaldehyde, and re-hydrated in PBS. After blocking for 45 min in BSA (in 5% PBS), the slides were incubated for 1 h with polyclonal rabbit anti-p-FAK and-paxillin antibodies (1∶50; Cell Signaling Technology), washed with PBS, and incubated for 45 min with Alexa 488-labeled goat anti-rabbit antibodies (1∶250; Molecular Probes). After rinsing in PBS, Hoechst 33258 was added for 15 min to counterstain the nuclei. The slides were washed with PBS and mounted with Vectashield (Vector Laboratories, Inc., Burlingame, CA, USA) then analyzed using fluorescence microscopy (Carl Zeiss).
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