Anti–mouse CD16/32 (FcR) mAb (2.4G2), anti–mouse CD11b mAb (5C6), control hamster Ig (UC8-1B9), anti–mouse CD4 mAb (GK1.5), anti–mouse CD8 mAb (53-6.7), anti–mouse FasL mAb (MFL3), and anti–mouse TRAIL mAb (N2B2) were prepared in our laboratory as previously described (4 (link), 19 (link)). Biotinylated anti–mouse F4/80 mAb (A3-1) was purchased from Caltag, biotinylated anti–mouse CD205 mAb (NLDC-145) was purchased from Cedarlane Laboratories, and biotinylated rat IgG2aκ (R35-95) and rat IgG 2bκ (A95-1) were purchased from BD Biosciences. A hamster IgG2 anti–mouse DR5 mAb (MD5-1) was generated by immunizing an Armenian hamster with mouse DR5-Ig fusion protein (DakoCytomation), fusing the splenocytes with P3U1 mouse myeloma cells, and then screening the reactivity to mouse DR5-transfected BHK (BHK-mDR5) cells by flow cytometry. MD5-1 was purified from ascites using a protein G column. Biotinylation was performed as previously described (20 (link)). Requests for MD5-1 should be addressed to H. Yagita (hyagita@med.juntendo.ac.jp ).
Protocol detail
Antibody Preparation and Handling
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A protein
Armenian hamster
Ascites
Biotinylation
Cd11b
Cd205
Cells
Fasl
Flow cytometry
Hamster
Igg2
Mouse
Myeloma
Protein
Protein g
Trail
Corresponding Organization :
Other organizations : Juntendo University, Peter MacCallum Cancer Centre, Science Applications International Corporation (United States), Frederick National Laboratory for Cancer Research, National Cancer Institute
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Protocol cited in 8 other protocols
Variable analysis
independent variables
- Anti–mouse CD16/32 (FcR) mAb (2.4G2)
- Anti–mouse CD11b mAb (5C6)
- Control hamster Ig (UC8-1B9)
- Anti–mouse CD4 mAb (GK1.5)
- Anti–mouse CD8 mAb (53-6.7)
- Anti–mouse FasL mAb (MFL3)
- Anti–mouse TRAIL mAb (N2B2)
- Biotinylated anti–mouse F4/80 mAb (A3-1)
- Biotinylated anti–mouse CD205 mAb (NLDC-145)
- Biotinylated rat IgG2aκ (R35-95)
- Biotinylated rat IgG 2bκ (A95-1)
- A hamster IgG2 anti–mouse DR5 mAb (MD5-1)
- Not explicitly mentioned
- Not explicitly mentioned
- Positive control: A hamster IgG2 anti–mouse DR5 mAb (MD5-1)
- Negative control: Control hamster Ig (UC8-1B9)
- Biotinylated rat IgG2aκ (R35-95) and biotinylated rat IgG 2bκ (A95-1) as additional controls
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