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In situ hybridization of a-satellite probes to human chromosomes 1, 15 and 17 detected by tyramide signal amplification. a-Satellite probes to chromosomes 1, 15 and 17 were labeled by nick translation with biotin-11-dUTP, ChromaTide® Texas Red®-12-dUTP (Prod # C7631) and ChromaTide® Oregon Green® 488-5-dUTP (Prod # C7630), respectively. Following simultaneous hybridization of all three probes, the biotinylated chromosome 1 probe was detected with HRP-streptavidin conjugate and Alexa Fluor® 546 tyramide (TSA Kit #23, Prod # T20933). HRP activity from this first TSA detection step was then quenched by treatment with 1% hydrogen peroxide for 30 minutes. Lastly, the Oregon Green® 488 dye-labeled chromosome 17 probe was detected with anti-fluorescein/Oregon Green® antibody (Prod # A6421) followed by HRP-conjugated goat anti-mouse IgG antibody and Alexa Fluor® 594 tyramide (TSA Kit #5, Prod # T20915). HRP activity from this second TSA detection step was then quenched by treatment with 1% hydrogen peroxide for 30 minutes. The Texas Red® dye-labeled chromosome 15 probe was then detected with rabbit anti-Texas Red® antibody (Prod # A6399) followed by HRP-conjugated goat anti-rabbit IgG antibody and Alexa Fluor® 488 tyramide (TSA Kit #12, Prod # T20922). After counterstaining with Hoechst 33258 (Prod # H1398, H3569, H21491), the images were acquired using filters appropriate for DAPI, FITC, TRITC and the Texas Red® dye.
|Tested species reactivity||Chemical|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||PBS, pH 7.2|
|Contains||5mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunohistochemistry (IHC)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Unlike biotin, which is an endogenous ligand in mitochondria, dye-based haptens permit background-free staining of cells and tissues and a sensitive alternative to biotin-streptavidin methods.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.