Activin A: Activin RIIB[Biotinylated] Inhibitor Screening Assay Kit
The Activin A: Activin RIIB [Biotinylated] Inhibitor Screening Assay Kit is an ELISA-based assay designed to measure the binding between Activin A and Activin RIIB (receptor IIB), also known as ACVR2B, protein for screening and profiling applications. This kit comes with enough purified Activin A (amino acids 311-426) and biotinylated Activin Receptor IIB (amino acids 19-137), streptavidin-HRP, assay buffer, and detection reagent for 100 enzyme reactions.
Figure 1. Activin A: Activin RIIB[Biotinylated] Inhibitor Screening Assay Kit schematic.
A 96-well plate is coated with Activin A protein. After coating and blocking, biotinylated Activin RIIB, also known as ACVR2B, is added in an optimized assay buffer. Next, unbound biotinylated Activin RIIB is washed away, and the plate is incubated with streptavidin-HRP. After a final wash, ELISA ECL substrate is added to produce chemiluminescence that can be measured using a chemiluminescence reader. The chemiluminescence signal is proportional to the efficacy of Activin RIIB binding to Activin A.
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- 1x PBS (Phosphate Buffer Saline) Buffer
- PBST Buffer (1x PBS, containing 0.05% Tween-20)
- Luminometer or microplate reader capable of reading chemiluminescence
- Adjustable micropipettor and sterile tips
- Rotating or rocker platform
| Catalog # | Name | Amount | Storage |
| Activin A | 1 µg | -80°C | |
| Activin RIIB, His-Tag, Avi-Tag, Biotin-Labeled* | <1 µg | -80°C | |
| 5x PP-02 Buffer | 4 ml | -80°C | |
| 79743 | Blocking Buffer 3 | 50 ml | -80°C |
| 79742 | Streptavidin HRP | 10 µl | -80°C |
| 79670 | ELISA ECL Substrate A (translucent bottle) | 6 ml | Room Temp |
| ELISA ECL Substrate B (brown bottle) | 6 ml | Room Temp | |
| 79837 | 96-well module plate | 1 | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
Activin A is a member of the TGFβ (transforming growth factor beta) family of proteins involved in embryonic development, hematopoiesis, cell proliferation, and cell differentiation. It is the ligand to the activin A receptor type I or type II, which are transmembrane receptors with serine/threonine kinase activity. Upon binding of activin, the kinase activity of the receptor is activated, SMAD2 (mothers against decapentaplegic homolog 2) and SMAD3 are phosphorylated, form a complex with SMAD4 and translocate to the nucleus, regulating gene expression. Activin A can be found in macrophages, dendritic cells, and neutrophils, playing a role in cell maturation and activation. It is involved in inflammation and autoimmune disorders, such as SLE (systemic lupus erythematosus), RA (rheumatoid arthritis), and atopic dermatitis. It is also involved in bone formation. The use of fusion protein blockers of the activin signaling pathways, that serve as sinks for activin A and other TGFβ members, is being explored for the treatment of pulmonary hypertension, chemotherapy-induced anemia, and osteoporosis.
Lodberg A., 2021 Cytokine & Growth Factor Reviews 60:1-17.
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