Thermo Fisher Scientific CD197 (CCR7) Monoclonal Antibody (4B12), APC-eFluor 780, eBioscience

Applications and Tested Dilution

• Application: Flow Cytometry (Flow)
• Tested Dilution: 1 µg/test
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Product Specifications

Available Formats

• Alexa Fluor 700
• APC
• Biotin
• eFluor 450
• Functional Grade
• PE
• PE-Cyanine7
• PE-eFluor 610
• PerCP-Cyanine5.5
• Request custom conjugation

Product Specific Information

The 4B12 monoclonal antibody reacts with mouse CCR7 (CD197), a chemokine receptor for CCL19 and CCL21. CCR7 mediates migration of naive T cells into secondary lymphoid organs (SLO) and plays a role in dendritic and B cell localization.
In humans, CCR7 marks central memory T cells (TCM) expressing CCR7 and CD62L. CCR7 deficiency leads to abnormal thymic development and impaired T cell maturation.

Applications

• Reported for flow cytometric analysis of mouse splenocytes.
• Tested at ≤1 µg/test (100 µL final volume; 10⁵–10⁸ cells/test).
• Optimize antibody titration for best performance.

Important Notes

• Staining requires specific conditions; 37°C incubation yields brighter and better-resolved staining than 2–8°C.
• Staining at 37°C does not interfere with other co-stains but should be validated per experiment.

Recommended Staining Protocol

1. Prepare cell suspension and block Fc receptors with 5 µg/million cells anti-mouse CD16/32 (Product #14-0161) for 15 min on ice.
2. Without washing, add 1 µg/million cells 4B12 antibody and incubate at 37°C (preferred) or 2–8°C for 30 min.
3. Wash cells once with 3 mL Flow Cytometry Staining Buffer (Product #00-4222) and decant.
4. Analyze by flow cytometry or proceed with secondary staining on ice.

Additional Notes

• Co-staining with CCL19-Fc may be limited due to binding condition differences and steric hindrance.
• 4B12 binding can block detectable CCL19-Fc binding.
• APC-eFluor® 780 is excited by the red laser (633 nm) and emits at 780 nm.
• Protect from light; tandem dye is photo-sensitive.
• Fixation compatible with IC Fixation Buffer (Product #00-8222) or 1-step Fix/Lyse Solution (Product #00-5333) for up to 3 days at 4°C.
• Filtration: 0.2 µm post-manufacturing filtered.

Target Information

CCR7 is a G protein-coupled receptor induced by Epstein-Barr virus (EBV) and expressed in blood, bone marrow, lymph node, and intestine. It regulates lymphocyte migration and dendritic cell maturation. Ligands include CCL19 (ECL) and CCL21.
CCR7 is crucial for T cell chemotaxis and phagocytic cell migration to inflammatory sites.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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