Human paraffin-embedded spleen tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate-Buffered Saline 1X (pH 6.0) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 (Cat. No. 925501) twice for 5 minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/ml of Alexa Fluor® 647 anti-CD45RA (clone HI100) antibody (green), Alexa Fluor® 594 anti-CD66b (clone G10F5) antibody (red) and Brilliant Violet 421™ anti-CD45RO antibody (blue) overnight at 4°C. The image was scanned with a 10X objective and stitched with MetaMorph® software.
Human paraffin-embedded spleen tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate-Buffered Saline 1X (pH 6.0) at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 (Cat. No. 925501) twice for 5 minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/ml of Alexa Fluor® 647 anti-CD45RA (clone HI100) antibody (green), Alexa Fluor® 594 anti-CD66b (clone G10F5) antibody (red) and Brilliant Violet 421™ anti-CD45RO antibody (blue) overnight at 4°C. The image was scanned with a 10X objective and stitched with MetaMorph® software.
CD66b is a 95-100 kD glycosylphosphatidylinositol (GPI)-linked protein also known as CD67, CGM6, and NCA-95. CD66b is a member of the immunoglobulin superfamily, carcinoembryonic antigen (CEA)-like subfamily. CD66b, expressed on granulocytes, has been reported to induce activation in neutrophils and to be involved in heterophilic adhesion with CD66c.
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions. The solution is free of unconjugated Alexa Fluor® 594.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 5.0 - 10.0 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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