NTB-A is a type I transmembrane protein belonging to the SLAM family of immune cell receptors and has been designated SLAMF6. The SLAM family proteins are members of the CD2 subgroup of the Ig superfamily. NTB-A is expressed on NK-, T-, and B cells. NTB-A exhibits homotypic interactions and can associate with adaptor molecules such as SAP to modulate immune cell functions.
Suitable for use in Flow Cytometry. Other applications not tested.
Flow Cytometry: 10ul labels 1x10e5 cells
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Cells should be Fc-blocked by treatment with 1ug of human IgG/10e5 cells for 15 minutes at RT prior to staining. Do not wash excess blocking IgG from this reaction.
2. Transfer 25ul of the Fc-blocked cells (1x10e5 cells) or 50ul of packed whole blood to a 5ml tube.
3. Add 10ul of N6000-35.
4. Incubate for 30-45 minutes at 4 degrees C.
5. Following this incubation, remove unreacted NTB- A/SLAMF6 reagent by washing the cells twice in 4ml of the same PBS buffer (Note: whole blood will require an RBC lysis step at this point using any commercially available lysing reagent).
6. Finally, resuspend the cells in 200-400ul of PBS buffer for final flow cytometric analysis.
7. As a control for analysis, cells in a separate tube should be treated with PE-labeled mouse IgG2a antibody.
Storage and Stability:
May be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C as an undiluted liquid. Dilute only prior to immediate use. Stable for 12 months at 4 degrees C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.