TNFSF13
Reactivity: Human, Mouse, Rat, Dog, Cow, Pig
WB, IHC, IF, IC
Host: Rabbit
Polyclonal
unconjugated
Application Notes
APRIL antibody can be used to recognize human APRIL by Flow Cytometry (also works for intracellular FC staining) and Western Blot. Anti-APRIL (MAb Aprily-1) does not cross react with mouse APRIL. Method: HEK 293 cells were mock transfected or transfected with an expression plasmid enabling surface expression of hAPRIL. Cells (5x105) were incubated on ice for 30 min in 50 μ,l FACS buffer (PBS, 5 % Fetal calf serum, 0.02 % azide) containing 10 μ,g/mL of Aprily-1 antibody. After washing in FACS buffer, FITC-conjugated antibody to mouse IgG was added. Cells were incubated on ice for 30 min, washed and analyzed by flow cytometry. Method: Cell supernatants were first precleared with protein-A sepharose and then subjected to immunoprecipitation with rhBCMA:Fc and protein-A sepharose. Beads were washed twice with PBS. Immunoprecipitated materials were eluted with 2.5 mM glycine pH 2.5 for 10 min, neutralized with Tris buffer and run on an SDS-PAGE gel and analyzed with 2 μ,g/mL Aprily-1 by Western blot. Note: In some migrations endogenous soluble APRIL resolved as a doublet around 20 kDa (dendritic cells: DC). rhsMegaAPRIL served as a positive control. Method: 3 HEK 293T cells (5 x 105) were mock transfected (thin line) or transfected with an expression plasmid enabling surface expression of mDR6 (thick line). Cells were incubated on ice for 30 min in 50 μ,l FACS buffer (PBS, 5 % Fetal calf serum, 0.02 % azide) containing 1 μ,g/mL of Luke-1 antibody. After washing in FACS buffer, PE-conjugated antibody to rat IgG was added. Cells were incubated on ice for 30 min, washed and then analyzed by flow cytometry.
Restrictions
For Research Use only
Format
Liquid
Buffer
PBS containing 0.02 % sodium azide
Preservative
Sodium azide
Precaution of Use
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
APRIL antibody can be stored at 4°C, stable for one year.
Target
TNFSF13
(Tumor Necrosis Factor (Ligand) Superfamily, Member 13 (TNFSF13))