In addition to flow cytometry analysis, canine CDCs were seeded onto fibronectin-coated chamber slides, after which the cells were fixed with 4% paraformaldehyde (PFA), blocked/permeabilized with Protein Block Solution (DAKO, Carpinteria, CA, USA) containing 1% saponin (Sigma-Aldrich), and then stained with anti-CD105 (ab156756; Abcam), anti-CD90 (mca1036g; AbD Serotec), anti-c-kit (ab5631; Abcam) and anti-CD45 (mca1042a488; AbD Serotec) antibodies. Fluorescein isothiocyanate (FITC)-secondary antibodies (Abcam) were used in conjunction with the aforementioned primary antibodies.
Fluorescein isothiocyanate fitc secondary antibodies
Manufactured by Abcam
Sourced in United Kingdom
Fluorescein isothiocyanate (FITC) secondary antibodies are a class of fluorescent-labeled antibodies used in various immunodetection techniques. FITC is a common fluorescent dye that conjugates to secondary antibodies, enabling the visualization of target proteins or molecules in samples.
Automatically generated - may contain errors
Lab products found in correlation
Protein block solution, by Agilent Technologies (2 mentions)
Saponin, by Merck Group (2 mentions)
Ab156756, by Abcam (1 mentions)
Ab5631, by Abcam (1 mentions)
Ab16669, by Abcam (1 mentions)
Mca48r, by Bio-Rad (1 mentions)
Epifluorescence microscope, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Tcl sl, by Leica (1 mentions)
3 protocols using fluorescein isothiocyanate fitc secondary antibodies
1
Canine CDC Phenotypic Characterization
2
Immunohistochemical Analysis of Cardiac Cell Therapy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rats were anesthetized with ketamine. Then, their hearts were exposed by left thoracotomy under sterile conditions. An MN-CSC patch containing 1 × 106 CSCs was placed onto the heart. Those rats that underwent the left thoracotomy but did not receive a patch were set as normal control. After 7 days, all rats were sacrificed, and hearts were collected and cryopreserved in optimum cutting temperature (OCT) compound. They were then cryosectioned and fixed with 4% paraformaldehyde. Protein block solution (Dako, Carpinteria, CA) containing 0.1% saponin (Sigma-Aldrich, St. Louis, MO) was used to permeabilize and protein-block each section. The following primary antibodies were used to target the desired proteins after an overnight incubation at 4°C: mouse anti-CD8α (1:100; MCA48R, AbD Serotec, Raleigh, NC) and rabbit anti-CD3 (1:100; ab16669, Abcam). Subsequently, fluorescein isothiocyanate (FITC) secondary antibodies (1:100; Abcam) were used for the detection of primary antibodies. DAPI (Life Technologies, NY, USA) was used to counterstain cell nuclei. Images were taken with an Olympus epifluorescence microscope.
3
Immunofluorescence Analysis of Cell-Cell Adhesion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were fixed with 3.7% paraformaldehyde followed by a permeabilization process using cold acetone. The cells were then blocked with 1% bovine serum albumin for 30 min, and incubated with N-cadherin or fibronectin antibody (Abcam, Cambridge, UK) at 1:300 dilutions in phosphate buffered saline (PBS) for 1 h at room temperature. The cells were then washed in PBS, incubated with fluorescein isothiocyanate (FITC) secondary antibodies (Abcam, Cambridge, UK) at a concentration of 1:600 dilution for 1 h in the dark, and counterstained for nuclei with Hoechst (Molecular Probes, Oregon, USA) for 10 min. Stained cells were analysed under a laser scanning confocal microscope (Leica TCL SL, Germany).
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!