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NDT401-ac
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.


Remarks:
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.

Reference:
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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