Cells were plated in confocal dishes and subjected to 8-Gy irradiation. After 48 h, the cells were fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.5% Triton for 20 min, blocked with 1% bovine serum albumin (BSA) for 1 h, and incubated with anti-GLI1 antibody (1:300, Abcam, Cambridge, MA, USA) at 4°C overnight and Cy3-labeled secondary antibody (1:100, goat anti-rabbit IgG, Boster, Wuhan, China). The dishes were scanned with a laser confocal scanning microscope (Zeiss LSM710, Carl Zeiss, Oberkochen, Germany).
Cy3 labeled secondary antibody
Manufactured by Boster Bio
Sourced in United States, China
The Cy3-labeled secondary antibody is a laboratory tool used for protein detection and visualization in various immunoassay techniques. It functions as a fluorescently-labeled detection reagent that binds to primary antibodies, allowing for the identification and localization of target proteins in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Beyotime (2 mentions)
Anti gli1 antibody, by Abcam (1 mentions)
Lsm 710, by Zeiss (1 mentions)
Alexa fluor 488 labeled secondary antibody, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Paraformaldehyde (pfa), by Sinopharm (1 mentions)
4 6 diamino 2 phenylindole dapi, by Beyotime (1 mentions)
Dapi staining solution, by Boster Bio (1 mentions)
Immunol staining wash buffer, by Beyotime (1 mentions)
Immunol staining primary antibody dilution buffer, by Beyotime (1 mentions)
Immnol fluorence staining secondary antibody dilution buffer, by Beyotime (1 mentions)
4 protocols using cy3 labeled secondary antibody
1
Quantifying GLI1 Expression in Irradiated Cells
2
Immunofluorescent Staining of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescent staining was conducted as described previously.52 (link) Briefly, at the indicated time points, cells were fixed with 4% paraformaldehyde for 15 min and permeabilized with 0.2% Triton X-100 (Beyotime, Shanghai, China) for 15 min followed by blocking with 10% goat serum (Beyotime, Shanghai, China) at room temperature for 1 h. Then, cells were incubated with the primary antibodies overnight at 4°C and then incubated with Cy3-labeled secondary antibody (BOSTER, Wuhan, China) or Alexa Fluor 488-labeled secondary antibody (Beyotime, Shanghai, China) for 1 h at 37°C in the dark. Subsequently, cells were stained with 4,6-diamidino-2-phenylindole (DAPI, Beyotime, Shanghai, China) for 10 min. Five random fields at 400 × magnification were photographed under a fluorescent microscope (Olympus BX53, Tokyo, Japan).
3
Immunofluorescent Identification of Rat NP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescence identification of primary rat NP cells was carried out by Procell. In brief, primary NP cells were fixed with 4% paraformaldehyde (Sinopharm, China) before permeabilization with 0.5% Triton X-100 (Beyotime, China). The cells were incubated with anti-type II collagen antibody (1:100, Cat No.: BA0533; Boster, China) at 4 °C overnight and then with Cy3-labeled secondary antibody (1:100, Cat No.: BA1032; Boster) at room temperature for 1 h. The nuclei were counterstained with 4,6-diamino-2-phenyl indole (DAPI) (Beyotime). A fluorescence microscope (BX53, Olympus, Japan) was used for image capture. The positive cells under five random high-power fields were counted under a light microscope, and the purity of NP cells was assessed by determining the type II collagen-positive cell rate.
4
Magnetic Stimulation Enhances VR-EPC Adhesion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VR-EPCs were seeded at a density of 1 × 104 cells/cm2 on the surfaces of Ti, TNrs, and Fe3O4-TNrs or into the plates wells without any sheets. VR-EPCs were cultured with M199 complete culture medium. The group with the addition of SEMF needed to be treated with 1 mT 15 Hz SEMF for 1 h per day. 3 days later, the cells were fixed with 4% paraformaldehyde for 15 min and then gently washed three times with Immunol Staining Wash Buffer (Beyotime, Shanghai, China). The cells were next treated with 1% BSA solution for 1 h, then incubated with NFATc1 primary antibody (ab25916, Abcam, MA, USA) diluted with Immunol Staining Primary Antibody Dilution Buffer (Beyotime, Shanghai, China) at 4 °C for 16 h. The cells were subsequently incubated at room temperature with Cy3 labeled secondary antibody (Boster, Wuhan, China) diluted with Immnol Fluorence Staining Secondary Antibody Dilution Buffer (Beyotime, Shanghai, China) for 1 h. After being gently washed, the cells were incubated with DAPI staining solution (Boster, Wuhan, China) for 5 min at room temperature. Then the cells could be observed and photographed under a fluorescence microscope.
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!