Primary chondrocytes were rinsed twice with PBS and fixed for 10 min at RT with 4% paraformaldehyde (PFA). Cells were rinsed rapidly twice with PBS and soaked in 0.2% Triton X-100 in PBS for 15 min at RT. Cells were then rinsed twice and soaked with Blocking One (Nacalai Tesque) for 1 h at RT, rinsed twice with PBS, and incubated with an anti-pStat3 (Tyr705) antibody (Cell Signaling) at 2 μg/mL in 5% Blocking One/PBS overnight at 4°C. The following day, cells were rinsed twice with PBS and incubated with Alexa Fluor 488 donkey anti-rabbit IgG (H+L) (Life Technologies) diluted to 1:1000 and Hoechst (Lonza) solution at 1 μg/mL in PBS for 1 h at RT.
Hoechst
Manufactured by Lonza
The Hoechst is a versatile lab equipment designed for a range of applications in scientific research and analysis. It serves as a specialized device for various tasks, including staining and fluorescence microscopy. The core function of the Hoechst is to enable precise and consistent sample preparation and analysis, but a detailed description of its intended use is not available.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 donkey anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
Blocking one, by Cell Signaling Technology (1 mentions)
Anti p stat3 tyr 705 antibody, by Cell Signaling Technology (1 mentions)
Blocking one, by Nacalai Tesque (1 mentions)
Af1924, by R&D Systems (1 mentions)
Rcab0003p, by ReproCELL (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Sc 5279, by Santa Cruz Biotechnology (1 mentions)
Mab4360, by Merck Group (1 mentions)
Mab4304, by Merck Group (1 mentions)
Alexa fluor 568, by Thermo Fisher Scientific (1 mentions)
Mab4303, by Merck Group (1 mentions)
Mab4381, by Merck Group (1 mentions)
N5413, by Merck Group (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
2 protocols using hoechst
1
Immunofluorescence Staining of Chondrocytes
2
Pluripotent Stem Cell Characterization
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ALP staining was performed with the ALP substrate (1-step NBT/BCIP; Pierce) after fixation with 4% paraformaldehyde (PFA; MUTO Pure Chemicals, Japan). Immunofluorescence staining was performed using the following primary antibodies: anti-NANOG (RCAB0003P, ReproCELL), anti-OCT3/4 (sc-5279, Santa Cruz), anti-SSEA 3 (MAB4303, Millipore), anti-SSEA 4 (MAB4304, Millipore), anti-Tra-1–60 (MAB4360, Millipore) and anti-Tra-1–81 (MAB4381, Millipore), anti-human smooth muscle actin (IR61161, DAKO), anti-human Sox17 (AF1924, R&D Systems), or anti-Nestin (N5413, Sigma). The fluorescence signals were detect using a conventional fluorescence laser microscope (IX70; Olympus) equipped with a color charge-coupled device (CCD) camera (CS220; Olympus). DAPI (Molecular Probes) and Hoechst (33342; Lonza) were used for nuclear staining. The secondary antibodies used were: anti-rabbit IgG and anti-mouse IgG and IgM conjugated with Alexa Fluor 488 or Alexa Fluor 568 (Molecular Probes).
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