Adult fly brains were dissected in cold PBS with 0.1% Triton-X (PBST) and fixed in 4% formaldehyde for 15–20 min on ice. Brains were rinsed 3 × 10 min with PBST, blocked for 30–60 min in 5% normal donkey serum in PBST (NDST), and incubated overnight (ON) at 4°C in primary antibody diluted in NDST. Brains were then rinsed 4 × 10 min in PBST, incubated 2 hr in secondary antibody diluted in NDST, rinsed 4 × 10 min in PBST, and mounted with Vectashield. The following primary antibodies were used: rabbit anti-GFP 1:1,000 (Molecular Probes A-11122), rabbit anti-DH44 1:500 (Johnson et al., 2005 (link)), guinea pig anti-PER 1:1,000 (I. Edery), mouse anti-PDF 1:500 (Developmental Studies Hybridoma Bank PDFC7; generated by J. Blau), and rabbit anti-SIFa 1:4,000 (gift from J. Veenstra). Secondary antibodies were as follows: Alexa Fluor 488 goat anti-rabbit 1:500 (Molecular Probes A-11008), fluorescein isothiocyanate (FITC) anti-guinea pig 1:500 (Jackson Laboratory 106-095-003), Cy5 donkey anti-mouse 1:400 (Jackson Laboratory 715-175-151), and Alexa Fluor 633 goat anti-rabbit (Molecular Probes A-21070). GFP signal from the CaLexA reporter system was detected with rabbit anti-GFP antibody followed by Alexa Fluor 488 goat anti-rabbit secondary antibody. Immunolabeled brains were visualized with a TCS SP5 confocal microscope.
Fluorescein isothiocyanate fitc anti guinea pig
Manufactured by Jackson ImmunoResearch
Fluorescein isothiocyanate (FITC) anti-guinea pig is a fluorescent labeling reagent used to detect and visualize guinea pig antigens in various biological applications, such as immunohistochemistry, flow cytometry, and immunofluorescence microscopy. It provides a bright green fluorescent signal when excited by a suitable light source.
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2 protocols using fluorescein isothiocyanate fitc anti guinea pig
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Immunohistochemistry of Drosophila Brains
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Immunohistochemistry of Drosophila Brains
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Adult fly brains were dissected in cold PBS with 0.1% Triton-X (PBST) and fixed in 4% formaldehyde for 15–20 min on ice. Brains were rinsed 3 × 10 min with PBST, blocked for 30–60 min in 5% normal donkey serum in PBST (NDST), and incubated overnight (ON) at 4°C in primary antibody diluted in NDST. Brains were then rinsed 4 × 10 min in PBST, incubated 2 hr in secondary antibody diluted in NDST, rinsed 4 × 10 min in PBST, and mounted with Vectashield. The following primary antibodies were used: rabbit anti-GFP 1:1,000 (Molecular Probes A-11122), rabbit anti-DH44 1:500 (Johnson et al., 2005 (link)), guinea pig anti-PER 1:1,000 (I. Edery), mouse anti-PDF 1:500 (Developmental Studies Hybridoma Bank PDFC7; generated by J. Blau), and rabbit anti-SIFa 1:4,000 (gift from J. Veenstra). Secondary antibodies were as follows: Alexa Fluor 488 goat anti-rabbit 1:500 (Molecular Probes A-11008), fluorescein isothiocyanate (FITC) anti-guinea pig 1:500 (Jackson Laboratory 106-095-003), Cy5 donkey anti-mouse 1:400 (Jackson Laboratory 715-175-151), and Alexa Fluor 633 goat anti-rabbit (Molecular Probes A-21070). GFP signal from the CaLexA reporter system was detected with rabbit anti-GFP antibody followed by Alexa Fluor 488 goat anti-rabbit secondary antibody. Immunolabeled brains were visualized with a TCS SP5 confocal microscope.
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