Activin A Protein, Human/Mouse/Rat (HEK293, His)

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Activin A, a multifunctional cytokine, is a member of TGF-β superfamily. Activin A first binds to the type II activin receptors (ActIIRA or ActRIIB) on the member surface, and then recruits and phosphorylates type I activin receptors (ActRI). Activin A primarily signal through SMAD2/3 proteins to regulate a variety of functions, including inflammation, fibrosis, and tumorigenesis. Activin A Protein, Mouse (HEK293, His) is produced in HEK293 cells with a C-Terminal His-tag.

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  • Species: Mouse
  • 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.
  • Biological Activity
  • Technical Parameters
  • Product Properties
  • Documentation
  • References
  • Help & FAQs

Biological Activity

Description

Activin A, a multifunctional cytokine, is a member of TGF-β superfamily. Activin A first binds to the type II activin receptors (ActIIRA or ActRIIB) on the member surface, and then recruits and phosphorylates type I activin receptors (ActRI). Activin A primarily signal through SMAD2/3 proteins to regulate a variety of functions, including inflammation, fibrosis, and tumorigenesis. Activin A Protein, Mouse (HEK293, His) is produced in HEK293 cells with a C-Terminal His-tag.

Background

Activin is a member of transforming growth factor-β (TGF-β) superfamily consisting of two inhibin β subunits linked by disulfide bonds. Activin A is expressed widely in various tissues and cells with strong bioactivities and is the mostly studied activin[1].
The sequence of amino acids in Activin A proteins from different species is very stable, which leads to the conclusion that in the process of evolution, Activin A has been only slightly altered, and that both in humans and in animals, its function is similar.
Activin exists in three basic molecular forms composed of two inhibin β subunits: activin A (βAβA), activin B (βBβB), and activin AB (βAβB). Activin A binds with high affinity to activin type II receptors, which recruit type I receptors and are necessary for the activation of Smad2/3 signaling. The phosphorylated ActRI activates Smad2 and Smad3, which form a complex with Smad4 to translocate to the nucleus. Activin and TGF-β share the same signaling pathway at the level of Smad2/3/4. Activin A exerts a variety of biological functions including regulation of hematopoietic cell proliferation, neuron differentiation, pituitary hormone secretion, and tissue repair. It is also involved in the process of many diseases, for example, inflammation, fibrosis and tumorigenesis[1][2].
Activin A has pro- and anti-tumorigenic functions depending on the tumor type. In breast, liver and colon cancers, activin signals were revealed to inhibit tumor cell growth. In addition, tumor tissues were revealed to express decreased levels of activin A or increased levels of activin antagonists or demonstrated the downregulation of activin receptors or Smad proteins. Moreover, its roles include embryonic differentiation, trophoblast invasion of the uterine wall in early pregnancy, and fetal/neonate brain protection in hypoxic conditions. Activin A also regulates bone formation and regeneration, enhances joint inflammation in rheumatoid arthritis, and triggers pathogenic mechanisms in the respiratory system[1][2].

In Vitro

Recombinant Activin A (-1 ng/mL; for 12 and 24 h) inhibits the proliferation of myeloma cell line NS-1 cells and induces NS-1 cell apoptosis. Activin A upregulates the expression of CHOP, GADD34, caspase-3, and caspase-12. Moreover, both Smad3 and p-Smad3 levels are increased with treatment of activin A in NS-1 cells[1].

In Vivo

Recombinant Activin A (2 ng/1 µL; intratumoral injection; for 6 days) inhibits the growth of solid tumors in tumor-bearing mice with NS-1 cells[1].

Verified Bioactivity

1.Measured in a cell proliferation assay using HT-29 human colon cancer cell. The ED50 for this effect is 6.407 ng/mL, corresponding to a specific activity is 1.561×105 units/mg.
2.Measured by its ability to inhibit proliferation of MPC-11 cells. The ED50 for this effect is typically ≤30 μg/mL.
3.Immobilized human ACVR2B at 10 μg/mL (100 μl/well) can bind biotinylated mouse INHBA-His, The EC50 of biotinylated mouse INHBA-His is ≤0.2 μg/mL.
4.Immobilized mouse INHBA-his at 10 μg/mL (100 μl/well) can bind human Follistatin Protein, The EC50 of human Follistatin Protein is ≤0.4 μg/mL.

MCE Validation Data

  • Purity - SDS-PAGE

    Purity - SDS-PAGE

    ≥ 90%, as determined by reducing SDS-PAGE.

  • Bioactivity - Cell-Based Assay

    Bioactivity - Cell-Based Assay

    Measured in a cell proliferation assay using HT-29 human colon cancer cell. The ED50 for this effect is 6.407 ng/mL, corresponding to a specific activity is 1.561×105 units/mg.

  • Bioactivity - Cell-Based Assay

    Bioactivity - Cell-Based Assay

    Measured by its ability to inhibit the proliferation of MPC-11 cells. The ED50 for this effect is typically 1.94 ng/mL, corresponding to a specific activity is 5.15×10^5 units/mg.

Technical Parameters

  • Species Mouse
  • Source HEK293
  • Tag C-6*His
  • Accession
  • Gene ID
  • Molecular Construction
    • N-term
    • Activin A (S21-S424)
      Accession # Q3UY39/NP_032406.1
    • 6*His
    • C-term
  • Protein Length

    Full Length of Mature Protein

  • Synonyms

    INHBA; Follicle-Stimulating Hormone-Releasing Protein; Inhibin Subunit Beta A; Erythroid Differentiation Factor; Inhibin Beta A Chain; Inhibin Beta A Subunit; Activin Beta-A Chain; FSH-Releasing Protein; Inhibin, Beta A (Activin A, Activin AB Alpha Polype

  • AA Sequence

    SPTPGSEGHGSAPDCPSCALATLPKDGPNSQPEMVEAVKKHILNMLHLKKRPDVTQPVPKAALLNAIRKLHVGKVGENGYVEIEDDIGRRAEMNELMEQTSEIITFAESGTARKTLHFEISKEGSDLSVVERAEVWLFLKVPKANRTRTKVTIRLFQQQKHPQGSLDTGDEAEEMGLKGERSELLLSEKVVDARKSTWHIFPVSSSIQRLLDQGKSSLDVRIACEQCQESGASLVLLGKKKKKEVDGDGKKKDGSDGGLEEEKEQSHRPFLMLQARQSEDHPHRRRRRGLECDGKVNICCKKQFFVSFKDIGWNDWIIAPSGYHANYCEGECPSHIAGTSGSSLSFHSTVINHYRMRGHSPFANLKSCCVPTKLRPMSMLYYDDGQNIIKKDIQNMIVEECGCS

  • Predicted Molecular Mass

    46.5 kDa

  • Molecular Weight

    Approximately 18 kDa (mature inhibin beta A chain) & 43 kDa (pro domain), based on SDS-PAGE under reducing conditions, due to the glycosylation.

  • Purity

    ≥ 90%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder

Formulation

1.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 5% trehalose, 5% mannitol, 0.01% Tween 80.
2.Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 150 mM NaCl, pH 7.4.
3.Lyophilized from a 0.22 μm filtered solution of PBS.
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.

References

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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