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Application: Useful for binding assays.
Assay Conditions: A white 96-well plate was precoated with APRIL, His-Avi-Tag Recombinant (#100254) (100 ng/well). Blocking Buffer 2 (#79728) and PBST were used for blocking and washing steps, respectively. For protein titration a serial dilution of Anti-APRIL Neutralizing Antibody (#102736) at the highest concentration of 10 ng/µl (250 ng/reaction) was prepared in 1x PP-02 Assay Buffer (#82620). Diluted Anti-APRIL Neutralizing Antibody was then added to the wells and plate was incubated at Room Temperature (RT) for 1 hour. The plate was then washed again and incubated with anti-Fc-HRP conjugate diluted 1:1000 in Blocking Buffer 2 for 1 hour at RT. After the final wash, ELISA ECL mix (#79670) was added to each well and chemiluminescence was read.
Background: APRIL (A proliferation-inducing ligand), also known as TNSF13 or CD256, activates a signaling pathway that participates in the regulation of B-cell development, autoimmunity, and long-term plasma cell survival. This protein is a member of the tumor necrosis factor (TNF) ligand family and a ligand for BCMA/ TNFRSF17. APRIL expression is significantly elevated in multiple myeloma (MM), and it has been demonstrated that activation of BCMA by APRIL promotes tumor growth, chemoresistance, and immunosuppression in the bone marrow microenvironment. Anti-APRIL monoclonal antibodies, such as sibeprenlimab and zigakibart, have recently progressed to clinical trials for Immunoglobulin A Nephropathy (IgAN) and MM, respectively. The development of a modified TACI (transmembrane activator and CAML interactor) domain resulted in Povetacicept, or ALPN-303, a dual inhibitor of APRIL and BAFF, with potential applications in the treatment of autoimmune disorders.
Description: This anti-APRIL neutralizing antibody is a purified, humanized recombinant monoclonal antibody that recognizes human APRIL (A proliferation-inducing ligand). This antibody has been tested for activity in a binding assay. It is similar to Sibeprenlimab.
Endotoxin Level: ≤3 EU/mg
Format: Aqueous buffer solution
Formulation: 8 mM phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
Host Cell Line: HEK293
Purity: ≥85%
Storage Stability: At least 6 months at -80°C.
Target: APRIL
Uniprot: O75888
Unit Definition: Antibody
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)