Human paraffin-embedded tonsil tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Critic buffer 1X pH 6.0 at 95°C for 40 minutes. Tissue was washed with PBS/ 0.05% Tween20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 5 µg/mL of purified anti-human CD8a (clone C8/144B) at 4°C overnight. On the next day, the tissue was washed twice with PBS and stained with rat anti-mouse IgG1 (clone RMG1-1) Alexa Fluor® 594 (red) and anti-human CD19 (clone A17136C) Alexa Fluor® 647 (green) for an hour at room temperature. The nuclei were counter staining with DAPI (blue). The image was scanned with a 10X objective and stitched with Metamorph® software.
Human paraffin-embedded tonsil tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Critic buffer 1X pH 6.0 at 95°C for 40 minutes. Tissue was washed with PBS/ 0.05% Tween20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 5 µg/mL of purified anti-human CD8a (clone C8/144B) at 4°C overnight. On the next day, the tissue was washed twice with PBS and stained with rat anti-mouse IgG1 (clone RMG1-1) Alexa Fluor® 594 (red) and anti-human CD19 (clone A17136C) Alexa Fluor® 647 (green) for an hour at room temperature. The nuclei were counter staining with DAPI (blue). The image was scanned with a 10X objective and stitched with Metamorph® software.
The RMG1-1 monoclonal antibody reacts with immunoglobulin G1 (IgG1) in all tested mouse haplotype (Igh-a and b). It does not react with other isotypes. The RMG1-1 monoclonal antibody may be used as a primary or secondary reagent for ELISA or immunofluorescent analysis.
Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 5 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
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