Primary and secondary antibodies used were as follows: rabbit polyclonal anti-phosphorylated myosin light chain 2 (Thr18/Ser19) (Cell Signaling Technology, #3674); Alexa 568 Fluor donkey anti-rabbit (ThermoFisher Scientific, A10042). Alexa Fluor 488 Phalloidin (A12379) and 4’,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI, D1306) were obtained from ThermoFisher. Y27632, Latrunculin A, Cytochalasin D, Calyculin A were purchased from Calbiochem. Rho Activator II was purchased from Cytoskeleton. Dulbecco’s modified Eagle’s medium (DMEM), DMEM/F12, and PBS were purchased from Gibco (Life Technologies); Fetal bovine serum (FBS) was purchased from Atlanta Biologicals; Epithelial growth factor (EGF) was purchased from Invitrogen; basic fibroblast growth factor (bFGF) was purchased from PeproTech; all other chemicals were from Sigma-Aldrich.
4 6 diamidino 2 phenylindole dihydrochloride dapi d1306
Manufactured by Thermo Fisher Scientific
Sourced in United States
4',6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI, D1306) is a fluorescent dye that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used as a nuclear counterstain in fluorescence microscopy to visualize cell nuclei.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 phalloidin a12379, by Thermo Fisher Scientific (2 mentions)
Rho activator 2, by Cytoskeleton (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Epithelial growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)
Ab101500, by Abcam (1 mentions)
Ab134035, by Abcam (1 mentions)
A11035, by Thermo Fisher Scientific (1 mentions)
2 protocols using 4 6 diamidino 2 phenylindole dihydrochloride dapi d1306
1
Antibody Detection of Cytoskeletal Dynamics
2
Immunofluorescence Staining of IL-22, CD4, and CD8
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Immunofluorescence was performed on 4-μm paraffin-embedded samples, as described elsewhere50 (link). After dewaxing, samples were fixed with paraformaldehyde 4% and blocked with phosphate-buffered saline with bovine serum albumin (PBS-BSA) 2% for 30 minutes. The primary antibodies IL-22 (ab134035, Abcam), CD4 (104R-16, Cell Marque, Rocklin, CA, USA), or CD8 (ab101500, Abcam) were incubated overnight at 4 °C. On the following day, the secondary antibodies, donkey anti-mouse Alexa 488 fluorescein (A21202, ThermoFisher Scientific, Waltham, MA, USA) (for IL-22 staining) and goat anti-rabbit Alexa 546 fluorescein (A11035, ThermoFisher Scientific) (for CD4 or CD8 staining), and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI, D1306, ThermoFisher Scientific) for nuclear identification were diluted and incubated for 90 minutes at room temperature. Images were acquired utilizing the appropriate filters of an Axiovert 200 inverted immunofluorescence microscope (Zeiss, Oberkochen, Germany).
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