Product Details
| Alternative Name: | A-Proliferation-inducing ligand, TNFSF 13, CD256 |
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| Clone: | Aprily-1 |
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| Host: | Mouse |
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| Isotype: | IgG1 |
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| Immunogen: | Recombinant human APRIL (aa 93-233). |
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| UniProt ID: | O75888 |
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| Source: | Purified from concentrated hybridoma tissue culture supernatant. |
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| Species reactivity: | Human
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| Crossreactivity: | Does not cross-react with mouse APRIL. |
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| Applications: | Flow Cytometry, IP, WB
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| Application Notes: | Flow Cytometry: also works for intracellular FC staining.
Excellent for Western Blot. |
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| Formulation: | Liquid. In PBS containing 0.02% sodium azide. |
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| Use/Stability: | Stable for at least 1 year after receipt when stored as recommended. |
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| Handling: | Avoid freeze/thaw cycles. |
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| Shipping: | Shipped on Blue Ice |
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| Short Term Storage: | +4°C |
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| Long Term Storage: | -20°C |
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| Regulatory Status: | RUO - Research Use Only |
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Figure 1: Western blot of total cell extracts from HEK 293 cells transfected with the indicated expression vector. Aprily-1 reacts specifically with human APRIL.Method: 10µg of protein was applied to the gel. Revealed with Aprily-1 (1µg/ml) and HRP-coupled anti-mouse secondary antibody (1:4’000).Note: The human APRIL construct used in this experiment is an uncleavable fusion protein between human BAFF (aa 1-132) and human APRIL (aa 93-233).
Figure 2: Detection of endogenous human APRIL with MAb to APRIL (Aprily-1) after IP with recombinant human BCMA:Fc.
Method: Cell supernatants were first precleared with protein-A sepharose and then subjected to immunoprecipitation with rhBCMA:Fc (Prod. No.
ALX-522-026) and protein-A sepharose. Beads were washed twice with PBS. Immunoprecipitated materials were eluted with 2.5mM glycine pH 2.5 for 10 min., neutralized with Tris buffer and run on an SDS-PAGE gel and analyzed with 2µg/ml Aprily-1 by Western blot.
Note: In some migrations endogenous soluble APRIL resolved as a doublet around 20kDa (dendritic cells: DC).
MegaAPRIL™, Soluble (human) (rec.) (Prod. No.
ALX-522-035) served as a positive control."
Figure 3:FACS analysis of cells with MAb to hAPRIL (Aprily-1).Method: HEK 293 cells were mock transfected or transfected with an expression plasmid enabling surface expression of a non-cleavable human APRIL. Cells (5x105) were incubated on ice for 30 min in 50 µl FACS buffer (PBS, 5% Fetal calf serum, 0.02% azide) containing 10µg/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.
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Product Literature References
BAFF production by antigen-presenting cells provides T cell co-stimulation: B. Huard, et al.; Int. Immunol.
16, 467 (2004),
Abstract;
General Literature References
BAFF and APRIL: A Tutorial on B Cell Survival: F. Mackay, et al.; Annu. Rev. Immunol. (2002),
Abstract;
