Cells were cultured on adhesive slides and treated with 20 µM ICG-001, 20 µM IQ-1, or vehicle alone for 24 h, fixed with prechilled methanol at −20 °C for 5 min and subsequently permeabilized with 0.2% Triton X-100 in 1 × PBS for 10 min. Immunostaining was carried out by using standard protocols. Primary antibodies were used at the following dilutions: anti-CK 17 (ab53707, 1:500) and anti-CK14 (sc-53253, Santa Cruz Biotechnology, 1:100). The secondary antibody was TRITC goat anti-mouse IgG (H+L) (ZF-0313, ZSGB-Bio, 1:100). After costaining with DAPI, cells were imaged by using an Olympus IX-71 microscope.
Sc 53253
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Sc-53253 is a laboratory product manufactured by Santa Cruz Biotechnology. It is a purified protein that can be used for research purposes. No further details about its core function or intended use can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Fitc and alexa 568 conjugated secondary antibodies, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Eclipse e600 microscope, by Zeiss (2 mentions)
Ix71 microscope, by Olympus (1 mentions)
Sc 74495, by Santa Cruz Biotechnology (1 mentions)
Ab 11333, by Abcam (1 mentions)
Ab150074, by Abcam (1 mentions)
5 protocols using sc 53253
1
Immunostaining of Cytokeratin Expression
2
Immunofluorescence Staining of Epithelial Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
3
Isolation and Characterization of Oral Cells
4
Immunostaining of Urothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
USCs or differentiated urothelia were fixed overnight in 10% formaldehyde at 4 °C. After washing in PBS, the fixed samples were pre-embedded into 2% agar, cut vertically, put in transwells side face up, embedded again in paraffin blocks and sectioned. The slides were stained for H&E and immunostained for selected antibodies. For immunofluorescence staining, slides were deparaffinized, hydrated, blocked with 10% heat-inactivated horse serum (HIHS) and 0.3% Triton X-100 in PBS, incubated with primary antibody in 1% HIHS and PBS overnight at 4 °C and secondary antibody in PBS for 30–60 min at room temperature6 (link). The primary antibodies used were mouse monoclonal anti-keratin 20 (Abcam, ab854, 1:200), goat polyclonal anti-E-cadherin (R&D Systems, AF748, 1:500), goat polyclonal anti-uroplakin 3a (Santa Cruz, sc-15186, 1:500), mouse monoclonal anti-uroplakin 3a (Fitzgerald, 10R-U103a, 1:50), rabbit polyclonal anti-p63 (GeneTex, GTX102425, 1:1,000), rabbit monoclonal anti-K5 (Abcam, ab150074, 1:100) and mouse monoclonal anti-keratin 14 (Santa Cruz, sc-53253, 1:50). Alexa Fluor secondary antibodies and DAPI were used at 1:1,000 dilution. Further antibody information is provided in Supplementary Table 3 . Samples were visualized on a Zeiss Axio Imager M2 Plus wide-field fluorescence microscope.
5
Immunofluorescence Staining of Epithelial Markers
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!