Cells were fixed with 100% methanol at −20°C for 10 min. Fixed samples were treated with blocking buffer (150 mM NaCl; 10 mM Tris-HCl, pH 7.5; 5% bovine serum albumin; and 0.1% Tween 20) for 30 min at 25°C, and incubated with primary and secondary antibodies overnight each at 4°C. Following each treatment, cells were washed 2–3 times with Dulbecco’s phosphate-buffered saline. Rat monoclonal anti-α-tubulin (YOL1/34, EMD Millipore); mouse monoclonal anti-centrin (20H5, EMD Millipore), goat anti-mouse Alexa Fluor 488-conjugated (ab150117, Abcam), and goat anti-rat Alexa Fluor 568-conjugated (ab175710, Abcam) antibodies were used at a dilution of 1:1000.
Ab175710
Manufactured by Abcam
Ab175710 is a recombinant monoclonal antibody fragment targeting a specific protein. This product is designed for use in various research applications, such as immunoassays and cellular analysis.
Automatically generated - may contain errors
3 protocols using ab175710
1
Immunofluorescence Labeling of Tubulin and Centrin
2
Cholangiocyte Characterization and Signaling
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Additional antibodies used for cholangiocyte characterization and signal transduction genes included: keratin 19 (K19, 1:10, Developmental Studies Hybridoma Bank, TROMAIII), collagen I (1:200; Southern Biotech, 1310-01), α-smooth muscle actin (mouse α-SMA, Abcam, ab7817). Secondary antibodies: PE (anti-mouse eBioscience, 12-4010-82, anti-rabbit #12-473981), APC (anti-mouse eBioscience, 17-4010-82). Alexa secondary antibodies- Abcam: goat anti-rabbit 647 (ab150083), goat anti-rat 568 (ab175710), goat anti-rabbit 555 (ab150086).
3
Immunofluorescence Staining of Microtubules
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Cells were fixed with 100% methanol at -20°C for 10 min. Fixed samples were treated with blocking buffer (150 mM NaCl; 10 mM Tris-HCl, pH 7.5; 5% bovine serum albumin; and 0.1% Tween 20) for 30 min at 25°C, and incubated with primary and secondary antibodies overnight each at 4°C. Following each treatment, cells were washed 2-3 times with Dulbecco's phosphate-buffered saline. Rat monoclonal anti--tubulin (YOL1/34, EMD Millipore); mouse monoclonal anti-centrin (20H5, EMD Millipore), goat anti-mouse Alexa Fluor 488-conjugated (ab150117, Abcam), and goat anti-rat Alexa Fluor 568-conjugated (ab175710, Abcam) antibodies were used at a dilution of 1:1000.
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