The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
Receptor involved in the costimulatory signal essential for T-lymphocyte proliferation and interleukin-2 production, by binding CD28 or CTLA-4. May play a critical role in the early events of T-cell activation and costimulation of naive T-cells, such as deciding between immunity and anergy that is made by T-cells within 24 hours after activation. Isoform 2 interferes with the formation of CD86 clusters, and thus acts as a negative regulator of T-cell activation.
Expressed by activated B-lymphocytes and monocytes.
All lanes : Anti-CD86 antibody (ab79683) at 1 µg/ml
Lane 1 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate Lane 2 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 4 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 6 : Human spleen tissue lysate - total protein (ab29699)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 37 kDa Observed band size: 37 kDa Additional bands at: 150 kDa, 35 kDa (possible cleavage fragment), 41 kDa (possible glycosylated form). We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
The band observed at 35 kDa represents the mature form of CD86 due to the presence of a 23 amino acid signal peptide.
The band observed at 41 kDa represents the pro-form of CD86 as it contains a number of potential glycosylation sites (SwissProt).