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a-Satellite probes to chromosomes 1, 15 and 17 were labeled by nick translation with biotin-11-dUTP, ChromaTide Texas Red-12-dUTP (C7631) and ChromaTide Oregon Green 488-5-dUTP, 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 Texas Red dyes.
|Tested species reactivity||Chem|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse|
|Storage buffer||0.1M sodium phosphate, pH 8, with 1.5% BSA|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
|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.
|Immunocytochemistry (ICC)||See 1 publications below|
Storage and reconstitution: a stock solution can be made by dissolving the powder in 0.5 mL of deionized water to yield an antibody concentration of 1 mg/mL. When stored desiccated at 4°C or -20°C, the lyophilized product is stable for at least six months. Stock solutions are stable for at least three months when stored at 4°C. For longer storage of solutions, divide into aliquots and freeze at -20°C.
Anti-fluorescent dye antibodies recognize specific fluorophores and, in most cases, quench their fluorescence. Thus many anti-dye antibodies, including those that recognize fluorescein, can serve as cell-impermeant probes for determining whether fluorescent dye-conjugated ligands, proteins, bacteria or other biomolecules have been internalized by endocytic or pinocytic processes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Ghrelin receptor as a novel imaging target for prostatic neoplasms.
A-6421 was used in immunocytochemistry to assess the specificity of a ghrelin-imaging probe for prostate cancer.
|Lu C,McFarland MS,Nesbitt RL,Williams AK,Chan S,Gomez-Lemus J,Autran-Gomez AM,Al-Zahrani A,Chin JL,Izawa JI,Luyt LG,Lewis JD||The Prostate (72:825)||2012|