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NDT401-ac Tissue preparation & colorimetric immunostaining with 2 primary antibodies on free-floating sections

This service includes tissue preparation,  sectioning, immunostaining, mounting, coverslipping and labeling the slides. As a result, you will receive up to 60 immunostained sections per brain or per tissue block ready for microscopic observations. To express our appreciation, a set of Nissl (or H & E) stained sections adjacent to those used for immunostaining will also be provided for you without additional cost.

Procedure:  Following cryoprotection, tissue will be rapidly frozen in isopentane pre-cooled to -70ºC.  The frozen tissue will then be cut on a cryostat and collected in our unique NDT301 Section cryopretection solution (NDT301). Subsequently, sections cut from various levels (or the levels of your choice) will be processed free-floating for immunohistochemistry with one specific antibody according to the avidin-biotin-complex (ABC) method1 (cf. photo samples below).

BrdUand NeuN double-immunostaining. 30 £gm cryostat section was cut from the hippocampal dentate gyrus of a mouse sacrificed 24 hr after the injection with 5-bromo-2-deoxyuridine (BrdU). The section was processed free-floating for BrdU-(blue) and then for NeuN-(red) immunoreactivity according to the ABC-AP method (cf. also photo samples of NDT401-bc).

C-fos and kir2.3 double immunostaining. 30 £gm cryostat section through the rat hypothalamus was processed free-floating for kir2.3- (brown) and then for c-fos- (red) immunoreactivities according to the avidin-biotin-complex method. Note nuclear labeling of c-fos immunoreactivity.

1. A quotation is required before placing an order.
2. The investigator needs to provide fixed tissue and the specific antibodies.
3. Please contact us for more information.

1. Hsu S. M., Raine L. and Fanger H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29, 577-580.

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