Mouse embryos were fixed at 4°C with Stefanini solution; 20 g/L Paraformaldehyde, 6.667% saturated picric acid and equal amounts of Sörensen Buffer 0.2M, pH7.2 and distilled water, followed by equilibration in 20% sucrose prior to 10 μm cryo-sectioning. Sections were permeabilized in PBS/0.1% Triton X-100, blocked with 10% goat or donkey serum (Sigma-Aldrich) and stained with primary antibodies: chicken anti-mouse eGFP (Millipore), rabbit anti-mouse MYL2 (Synaptic Systems), rat anti-mouse ITGA6 Allophycocyanin (APC) (eBioscience), rat anti-mouse ITGA5 Biotin (LifeSpan Biosciences) and rabbit anti-mouse cTropT PE (BD Biosciences). Primary antibodies were visualized with secondary antibodies conjugated to FITC/Cy3/Cy5 (Jackson ImmunoResearch) or Alexa Fluor 555/647 (Invitrogen). Hoechst 33342 (Invitrogen) was used to localize nuclei. Imaging was performed using a Zeiss LSM 780 (Germany) laser scanning confocal microscope or a Leica DM500 B (Switzerland).
Fitc cy3 cy5
Manufactured by Jackson ImmunoResearch
FITC/Cy3/Cy5 is a set of fluorescent dyes commonly used in immunoassays and other biological applications. FITC (Fluorescein Isothiocyanate) is a green fluorescent dye, Cy3 is an orange-red fluorescent dye, and Cy5 is a far-red fluorescent dye. These dyes can be conjugated to various biomolecules, such as antibodies, proteins, or nucleic acids, to facilitate their detection and visualization in experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780, by Zeiss (2 mentions)
Goat or donkey serum, by Merck Group (1 mentions)
Rat anti mouse itga5 biotin, by LifeSpan BioSciences (1 mentions)
Rabbit anti mouse ctropt pe, by BD (1 mentions)
Alexa fluor 555 647, by Thermo Fisher Scientific (1 mentions)
Dm500b, by Leica camera (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Gh2ax, by Abcam (1 mentions)
Sycp1, by Abcam (1 mentions)
Elyra ps 1 microscope system, by Zeiss (1 mentions)
Sycp3, by Abcam (1 mentions)
Lsm780 meta inverted confocal microscope, by Zeiss (1 mentions)
Rad51, by Santa Cruz Biotechnology (1 mentions)
2 protocols using fitc cy3 cy5
1
Multiparametric Embryo Immunohistochemistry
2
Immunofluorescence Analysis of Meiotic Proteins
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Cultured cells were digested into single-cell suspensions. Chromosomal spreads were prepared using a hypotonic bursting technique (Peters et al., 1997) . Primary antibodies were Sycp3 (Abcam), Sycp1 (Abcam), gH2AX (Abcam), Rad 51 (Santa Cruz), and Spo11 (provided by Scott Keeney) (Lange et al., 2011) . Secondary antibodies were FITC-, Cy3-, Cy5-, and DyLight 405labeled (Jackson ImmunoResearch). Images were captured with Zeiss LSM780 Meta inverted confocal microscope. Super-resolution analysis was performed using a Zeiss Elyra PS.1 microscope system.
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