Multi-color FISH was performed as previously described in (35 (link)) with minor modifications. Tissues were serially rehydrated, and the pituitary was detached from the brain. Afterwards, whole pituitaries were hybridized with the probes (0.11 – 3.17 ng/µl) for 18 hours at 55°C, and incubated with different combinations of anti-DNP- (Perkin Elmer), anti-FITC-, and anti-DIG-conjugated antibodies (Roche Diagnostics), followed by TAMRA- (Thermofisher), Cy5- (Perkin Elmer) and FITC-conjugated tyramides (Sigma). The nuclei were stained with DAPI (1:1000, 4’, 6-diamidino-2-phenylindole dihydrochloride; Sigma). The absence of labeling when using sense probes was used to confirm the specificity of the anti-sense probes. Whole pituitaries were mounted using Vectashield H-1000 Mounting Medium (Vector, Eurobio/Abcys) between microscope slides and cover slips (Menzel Glässer, VWR) with spacers (Reinforcement rings, Herma) in between for the juveniles, and between two cover slips with spacers for adults.
Anti fitc
Manufactured by Roche
Sourced in Switzerland
The Anti-FITC- is a laboratory equipment product designed for specific applications. It serves a core function in certain experimental procedures, but a detailed description without interpretation or extrapolation cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Tamra, by Thermo Fisher Scientific (2 mentions)
Anti dig conjugated antibodies, by Roche (2 mentions)
Anti dnp, by PerkinElmer (2 mentions)
Fitc conjugated tyramides, by Merck Group (2 mentions)
Vectashield mounting medium h 1000, by Vector Laboratories (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
H 1000 mounting medium, by Vector Laboratories (1 mentions)
Fitc dutp, by Roche (1 mentions)
3 protocols using anti fitc
1
Multi-color FISH Assay for Pituitary Cells
2
Multi-color FISH Pituitary Analysis
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Multi-color FISH was performed as previously described in (34) with minor modifications. Tissues were serially rehydrated, and the pituitary was detached from the brain. Afterwards, whole pituitaries were hybridized with the probes (0.11 -3.17 ng/µl) for 18 hours at 55 o C, and incubated with different combinations of anti-DNP-(Perkin Elmer), anti-FITC-, and anti-DIG-conjugated antibodies (Roche Diagnostics), followed by TAMRA-(Thermofisher), Cy5-(Perkin Elmer) and FITC-conjugated tyramides (Sigma). The nuclei were stained with DAPI (1:1000, 4', 6diamidino-2-phenylindole dihydrochloride; Sigma). The absence of labeling when using sense probes was used to confirm the specificity of the anti-sense probes. Whole pituitaries were mounted using Vectashield H-1000 Mounting Medium (Vector, Eurobio/Abcys) between microscope slides and cover slips (Menzel Glässer, VWR) with spacers (Reinforcement rings, Herma) in between for the juveniles, and between two cover slips with spacers for adults.
3
Quantifying Kidney Apoptosis via TUNEL
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We used the method of TUNEL (Roche Co., Basel, Switzerland) to assess the apoptosis of kidney tubular epithelial cells (KTEC). The kidneys were embedded in paraffin, sliced (2 μm), and dewaxed in xylene. Proteinase K was added, and the mixture was incubated for 12 min at room temperature. The sections were washed, TdT, FITC-dUTP, and FITC-dATP (Roche Co., Basel, Switzerland) added gradually, incubated for one hour in a humidified incubator, anti-FITC (Roche Co., Basel, Switzerland) added, and DAB stained. The number of apoptotic cells was observed under a 400-fold magnification under a light microscope, and three fields were randomly selected from each slice. We calculated the apoptotic rate of KTEC by dividing the positive cells by the total number of cells.
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