ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His)

ACVR2A, also known as Activin RIIA, is an activin type II receptor. On ligand binding, ACVR2A can forms a receptor complex consisting of two type II and two type I transmembrane serine/threonine kinases. ACVR2A is a receptor for Activin A, Activin B and Inhibin A^[1][2]. ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His) is produced in HEK293 cells with six C-Terminal His-tags. It consists of 115 amino acids (A20-P134).

ACVR2A is a type II member of the TGF-β family of receptor Serine/Threonine kinases. ACVR2A is a receptor for Activin A, Activin B and Inhibin A^[1][2].
The sequence of amino acids in ACVR2A proteins from different species is very stable, which leads to the conclusion that in the process of evolution, ACVR2A has been only slightly altered, and that both in humans and in animals, its function is similar.
Signaling by activins and BMPs is highly promiscuous, since apart from signaling through ALK4/7, the activin type II receptors (ACVR2A and 2B) can interact also with several type I BMP receptors (ALK1/2/3/6), which can also form complexes with the type II BMP receptor, BMPRII. Type II receptors phosphorylate and activate type I receptors which autophosphorylate, then bind and activate SMAD transcriptional regulators. ACVR2A can form complexes with different type I receptors that signal either to Smad2/3 (ALK4) or to Smad1/5/8 (ALK2, ALK3, ALK6). The different type I receptors compete for binding to ACVR2A and that this competition provides a mechanism that balances signaling between Activin A-mediated, ALK4-dependent Smad2/3 signaling, and BMP-mediated ALK2 or ALK3-dependent signaling to Smad1/5/8. In myeloma cells, BMP-6- and BMP-9-induced activation of SMAD1/5/8 through ACVR2A/ACVR2B/ALK2 is inhibited by activin A treatment^[1][3].

Recombinant human ACVR2A (5 μg/mL) inhibits the effects of BMP-9 on INA-6 and IH-1 cells^[1].
Recombinant human ACVR2A (100 μg/mL; 96 hours) blocks the β-cell-proliferative effect of adenoviruses containing mouse Pdx-1 (AdCMV-mPdx-1) treatment^[4].

1.This product does not contain protein kinase domain.
2.Measured by its ability to neutralize Activin A inhibitory effect of murine MPC-11 cells. The ED₅₀ this effect is 0.0487 μg/mL in the presence of 7.5 ng/mL Activin A, corresponding to a specific activity is 2.05×10⁴ units/mg.
3.Measured by its ability to neutralize Activin-mediated inhibition on MPC-11 cell proliferation. The ED₅₀ for this effect is <0.1 μg/mL in the presence of 10 ng/ml Recombinant Human Activin A.

• 
  Measured by its ability to neutralize Activin A inhibitory effect of murine MPC-11 cells. The ED₅₀ for this effect is 0.0487 μg/mL in the presence of 7.5 ng/mL Activin A, corresponding to a specific activity is 2.05×10⁴ units/mg.

Cell Culture
Cell Line for Activity Assay: MPC-11 cells
Cell Culture Medium: DMEM + 10% Horse Serum (HS) + 1% Penicillin/Streptomycin (P/S)
Test protein: Activin A Protein, Human/Mouse/Rat (HEK293) (HY-P70311)
Penicillin-Streptomycin (100×), Sterile（HY-K1006）
Cell Counting Kit-8 (HY-K0301)

Procedure
1. Seed cells at a density of 15,000 cells per well in a 96-well plate, with a volume of 50 μL per well. (Do not seed cells in the outermost ring of the 96-well plate; instead, add an equal volume of PBS solution.)
2. Add 50 μL of complete medium (containing 10 ng/ml Activin A) with different concentrations of ACVR2A protein to the 96-well plate, resulting in final concentrations of 0, 0.01, 0.1, 0.5, 1, 5, 10, 50, 100, 500, 1000, 5000, and 10000 ng/mL of ACVR2A protein in each well.
3. Incubate the cells at 37°C in a 5% CO₂ incubator for 72 hours.
4. After incubation, add 10 μL of CCK8 solution to each well under light-protected conditions and incubate at 37°C in a 5% CO₂ incubator for 3 hours.
5. Measure the OD value at 450 nm using a microplate reader.
6. Using GraphPad Prism software, plot a curve with OD values on the y-axis and protein concentration on the x-axis to calculate the ED₅₀ value.

Cynomolgus; Human

HEK293

C-6*His

P27037-1/G7PKJ4 (A20-P134)

92  [NCBI]/102121129

Extracellular Domain

AILGRSETQECLFFNANWEKDRTNQTGVEPCYGDKDKRRHCFATWKNISGSIEIVKQGCWLDDINCYDRTDCVEKKDSPEVYFCCCEGNMCNEKFSYFPEMEVTQPTSNPVTPKP

Approximately 24-40 kDa due to the glycosylation.

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  ≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 150 mM NaCl, 1 mM DTT, pH 7.4 or PBS, pH 7.4, 8% trehalose.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.

• [1]. /biology-dictionary/snb-19-cell.html  [Content Brief]