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

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Based on 1 Customer Validation

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. 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).

For research use only. We do not sell to patients.
  • Species: Cynomolgus; Human
  • Source: HEK293
  • Storage:
    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.
  • Experimental Validation Results for ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His)
  • Experimental Validation Results for ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His)
  • ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His) Product Image
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  • Biological Activity
  • Technical Parameters
  • Product Properties
  • Documentation
  • Help & FAQs
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Biological Activity

Description

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).

Background

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].

In Vitro

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].

Verified Bioactivity

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.

MCE Validation Data

  • Purity - SDS-PAGE

    Purity - SDS-PAGE

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

  • Bioactivity - Cell-Based Assay

    Bioactivity - Cell-Based Assay

    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.

Assay Procedure

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.

Technical Parameters

  • Species Cynomolgus; Human
  • Source HEK293
  • Tag C-6*His
  • Accession

    P27037-1/G7PKJ4 (A20-P134)

  • Gene ID
    92  [NCBI]/102121129
  • Molecular Construction
    • N-term
    • ACVR2A (A20-P134)
      Accession # P27037-1
    • 6*His
    • C-term
  • Protein Length

    Extracellular Domain

  • Synonyms

    ACVR2A; Activin Receptor Type-2A; Prev. ACVR2; Activin Receptor Type IIA; ACTRII; ACTR-IIA; Activin A Receptor, Type IIA; ACTRIIA; Activin A Receptor Type 2A; Activin A Receptor, Type II

  • AA Sequence

    AILGRSETQECLFFNANWEKDRTNQTGVEPCYGDKDKRRHCFATWKNISGSIEIVKQGCWLDDINCYDRTDCVEKKDSPEVYFCCCEGNMCNEKFSYFPEMEVTQPTSNPVTPKP

  • Predicted Molecular Mass

    14.1 kDa

  • Molecular Weight

    Approximately 26-40 kDa, based on SDS-PAGE under reducing conditions.

  • Glycosylation

    Yes

  • Purity

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

Product Properties

Appearance

Lyophilized powder.

Formulation

1.Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 150 mM NaCl, 1 mM DTT, pH 7.4.
2.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
Please refer to the lot-specific COA for specific buffer information.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

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).

Storage & Stability

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.

Shipping

Room temperature in continental US; may vary elsewhere.

Calculators

Reconstitution Calculator

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

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Dilution Calculator

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
Unit/mg
= 106 ÷
Biological Activity (ED50) Biological Activity (ED50)
106 ÷
ng/mL
MOQ
Minimum order quantity
100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

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

  • 1/16

    MedChemExpress Validation
     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.

  • ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His) Product Image
    1/16
  • Experimental Validation Results for ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His)
  • Experimental Validation Results for ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His)
  • ACVR2A/Activin RIIA Protein, Human/Cynomolgus (HEK293, His) Product Image