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Moab a-Bovine adenovirus 3


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Descripción

I – INDIRECT IMMUNOFLUORESCENCE ASSAY PROCEDURE
Fix the cell preparation (cell culture or tissue sections) for 15 minutes at 21°C +/- 3°C using one of the
fixatives listed below:
– 2% paraformaldehyde in PBS
– Acetone (9 V) + Water (1 V)
– Ethanol (1 V) – Acetone (1 V)
– Pure isopropanol solution
– Absolute ethanol solution
Then rinse with PBS.
Dilute the reagent twentyfold in PBS prepared according to the following formula:
Phosphate Buffer Saline (PBS)
___________________________
NaCl: 8 gm
KH2PO4: 0.2 gm
KCl: 0.2 gm
Na2HPO4 . 2H2O: 1.15 gm
NaN3: 0.1 gm
H20 1 L
Incubate the preparation on the sample for 1 hour at 21°C +/- 3°C, preferably in a humidity chamber.
Upon completion of this incubation period rinse the preparation with a PBS solution.
Then add the conjugate (fluorescein-labelled anti-mouse immunoglobulin) at the manufacturer’s
recommended dilution. The conjugate available from Bio-X Diagnostics (BIO 156) should be diluted
twentyfold in PBS-Evans Blue solution.
PBS-Evan Blue
______________
PBS 1 liter
Evans Blue: 0.01 gm
Incubate the preparation on the sample for 1 hour at 21°C +/- 3°C, preferably, in a humidity chamber. After this second incubation step rinse the preparation with PBS.

Dry the slide, then add the mounting medium made up as follows:
Mounting medium
_________________
Glycerol 9 parts by volume
PBS 1 part by volume
Place a cover slip on the slide, then observe under a microscope fitted for fluorescence detection.
The antibody may be kept in its original vial between +2°C and +8°C for more than a year.
Never freeze this reagent.
Once diluted in PBS, the antibody remains stable for one week between +2°C and +8°C.
II – INDIRECT IMMUNOPEROXIDASE ASSAY PROCEDURE
Fix the cell preparation (cell culture or tissue sections) for 15 minutes at 21°C +/- 3°C using one of the
following fixatives:
– 2% paraformaldehyde in PBS
– Acetone (9 V) + Water (1 V)
– Ethanol (1 V) – Acetone (1 V)
– Pure isopropanol solution
– Absolute ethanol solution
Then rinse with PBS.
Dilute the reagent twentyfold in PBS prepared according to the following formula:
PBS
______
NaCl: 8 gm
KH2PO4: 0.2 gm
KCl: 0.2 gm
Na2HPO4 . 2H2O: 1.15 gm
NaN3: 0.1 gm
H20 1 L
Incubate the preparation on the sample for 1 hour at 21°C +/- 3°C, preferably in a humidity chamber.
Upon completion of this incubation period rinse the preparation with PBS.
Then add the conjugate (peroxidase-coupled anti-mouse immunoglobulin) at the manufacturer’s recommended
dilution. The conjugate available from Bio-X Diagnostics (BIO 157) should be diluted twentyfold in PBS.
Incubate the preparation on the sample for 1 hour at 21°C +/- 3°C, preferably in a humidity chamber.
After this second incubation step rinse the preparation with PBS.
Then add the chromogen (AEC, precipitating TMB, DAB, etc.) and the substrate (hydrogen peroxide)
according to the manufacturer’s instructions. Examine under the microscope for the presence of the coloured
marker.

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