Activin A betaA subunit Antibody (69403)

R&D Systems | Catalog # MAB3381

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Porcine

Applications

Validated:

ELISA Capture (Matched Antibody Pair), Neutralization

Cited:

Immunohistochemistry, Western Blot, Neutralization, Immunocytochemistry, Bioassay, Blocking, ELISA Capture, ELISA Development, In vivo assay, Mass Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 69403
View all Activin A betaA subunit Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human Activin A

Specificity

Detects recombinant human Activin A in ELISAs. Also recognizes Activin A precursor. The amino acid sequences of mature beta A subunits from human, mouse and rat are identical. Therefore, this antibody will also detect the mature beta A subunit from mouse and rat.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Activin A betaA subunit Antibody (69403)

Hemoglobin Expression Induced by Activin A and Neutralization by Human/ Mouse/Rat Activin A Antibody.

Hemoglobin Expression Induced by Activin A and Neutralization by Human/ Mouse/Rat Activin A Antibody.

Recombinant Human/Mouse/ Rat Activin A (Catalog # 338-AC) increases hemoglobin expression in the K562 human chronic myelogenous leukemia cell line in a dose-dependent manner (orange line), as measured by the psuedoperoxidase assay. Hemoglobin Expression elicited by Recombinant Human/Mouse/Rat Activin A (7.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Mouse/Rat Activin A beta A subunit Monoclonal Antibody (Catalog # MAB3381). The ND50 is typically 0.02-0.06 µg/mL.
Detection of Activin A beta A subunit by Immunohistochemistry

Detection of Activin A beta A subunit by Immunohistochemistry

Activin A levels are significantly higher in tumor tissue and correlate to worse prognosis in PDAC. (A) Representative cores of pancreatic tumors from a human pancreatic tissue microarray stained with H&E to assess the percentage of epithelial versus stromal cells in each core. Adjacent sections were immunostained for activin A expression. (B) Quantification of the percentage of epithelial cells (left panel) or stromal cells (right panel) in normal tissue (white bar) versus tumor tissue (black bar). Statistical analysis is unpaired t-test. (C) Quantification of activin A in TMA (n = 63). An average of the percentage of epithelial and stromal fraction was calculated and scored in each TMA. Comparing epithelial (left panel) and stromal fraction (right panel) in normal tissue (white bar) versus tumor tissue (black bar). Statistical analysis 2-way ANOVA; mean ± SEM, ***(p < 0.001). (D) Kaplan–Meier plots of patient overall survival versus expression of activin A in either epithelial cells (left panel) or stromal cells (right panel). Statistical analysis Log-rank (Mantel-Cox) test, ** (p < 0.01), ***(p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33846512), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Immunohistochemistry

Detection of Activin A beta A subunit by Immunohistochemistry

Inhibition of activin A in KPC mice significantly reduces metastasis. (A) Time course of anti-activin A neutralizing antibody injected in mice weekly (2 mg/kg). Control animals were injected with the same concentration of IgG isotype. Remaining animals were collected at the end of week 13 of treatment. (B) Representative image (×4 magnification) of H&E staining of pancreas from control treated (IgG) and anti-activin A treated KPC mice IHC-stained for either alpha -SMA (to indicate stromal cells) or activin A. Representative stromal cells marked by arrows. (C) Representative image (×4 magnification) of H&E staining of pancreas used for quantification of CN (yellow arrow) cystic lesions. Quantification of lesions is documented in both IgG control and anti-activin A antibody treated mice. (D) Left panel: Representative images (×10 and ×40 magnification) of liver tissue from control treated versus anti-activin A treated KPC mice stained with H&E (upper panel) or ductal marker protein CK19 (Lower panel). Right panel: Quantification of distant metastatic formation in the liver tissue of control (n = 5) and treated (n = 7) mice. Statistical analysis unpaired Student t-test (ns: not significant). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33846512), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Immunohistochemistry

Detection of Activin A beta A subunit by Immunohistochemistry

Inhibition of activin A in KPC mice significantly reduces metastasis. (A) Time course of anti-activin A neutralizing antibody injected in mice weekly (2 mg/kg). Control animals were injected with the same concentration of IgG isotype. Remaining animals were collected at the end of week 13 of treatment. (B) Representative image (×4 magnification) of H&E staining of pancreas from control treated (IgG) and anti-activin A treated KPC mice IHC-stained for either alpha -SMA (to indicate stromal cells) or activin A. Representative stromal cells marked by arrows. (C) Representative image (×4 magnification) of H&E staining of pancreas used for quantification of CN (yellow arrow) cystic lesions. Quantification of lesions is documented in both IgG control and anti-activin A antibody treated mice. (D) Left panel: Representative images (×10 and ×40 magnification) of liver tissue from control treated versus anti-activin A treated KPC mice stained with H&E (upper panel) or ductal marker protein CK19 (Lower panel). Right panel: Quantification of distant metastatic formation in the liver tissue of control (n = 5) and treated (n = 7) mice. Statistical analysis unpaired Student t-test (ns: not significant). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33846512), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Western Blot

Detection of Activin A beta A subunit by Western Blot

Specific activin A inhibition attenuates TGF beta 1-induced Smad3 activation and profibrotic responses in MC. a ELISA demonstrates that TGF beta 1 (24 h) increases actA and actB secretion (n = 3) to 19.5 ng/ml and 2.5 ng/ml, representing an 8.9- and 1.06-fold induction respectively. b TGF beta 1 increases actA in whole cell lysate by 1.8-fold (n = 3). c ActA (20 ng/ml) upregulates FN (n = 3–4), CTGF (n = 4) and alpha SMA (n = 6) at 48 h. d TGF beta 1 and actA both increase Smad3 transcriptional activity; no synergistic effect is seen (n = 6–12). e An actA neutralizing antibody attenuates TGF beta 1-induced FN (n = 5–6), alpha SMA (n = 5), CTGF (n = 5–6), and Smad3 activation (n = 10–12). f ActA neutralization decreases TGF beta 1-induced Smad3 transcriptional activity at 24 h (n = 9–15), but this is not decreased by actB neutralization (n = 6) (g). h MC were stimulated with TGF beta 1 or actA for 1 h, then treated with their type I receptor inhibitor SB431542 (50 µM). Restimulation with the same ligand shows that cells become refractory to TGF beta 1, but not actA (n = 4). *, **, ***, ****P < 0.05, 0.01, 0.001, 0.0001; one-way ANOVA with Tukey’s multiple comparisons post hoc test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36717814), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Western Blot

Detection of Activin A beta A subunit by Western Blot

Specific activin A inhibition attenuates TGF beta 1-induced Smad3 activation and profibrotic responses in MC. a ELISA demonstrates that TGF beta 1 (24 h) increases actA and actB secretion (n = 3) to 19.5 ng/ml and 2.5 ng/ml, representing an 8.9- and 1.06-fold induction respectively. b TGF beta 1 increases actA in whole cell lysate by 1.8-fold (n = 3). c ActA (20 ng/ml) upregulates FN (n = 3–4), CTGF (n = 4) and alpha SMA (n = 6) at 48 h. d TGF beta 1 and actA both increase Smad3 transcriptional activity; no synergistic effect is seen (n = 6–12). e An actA neutralizing antibody attenuates TGF beta 1-induced FN (n = 5–6), alpha SMA (n = 5), CTGF (n = 5–6), and Smad3 activation (n = 10–12). f ActA neutralization decreases TGF beta 1-induced Smad3 transcriptional activity at 24 h (n = 9–15), but this is not decreased by actB neutralization (n = 6) (g). h MC were stimulated with TGF beta 1 or actA for 1 h, then treated with their type I receptor inhibitor SB431542 (50 µM). Restimulation with the same ligand shows that cells become refractory to TGF beta 1, but not actA (n = 4). *, **, ***, ****P < 0.05, 0.01, 0.001, 0.0001; one-way ANOVA with Tukey’s multiple comparisons post hoc test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36717814), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Immunohistochemistry

Detection of Activin A beta A subunit by Immunohistochemistry

Activin A neutralization inhibits renal fibrosis in TGF beta 1-overexpressing mice. a TGF beta 1 transcript is increased in mice genetically engineered to overexpress TGF beta 1 (HH) compared with wild-type mice (WT) (n = 6–7, *p ≤ 0.05). b Serum actA is elevated in wild-type and HH mice after UUO. This is decreased by treatment with a neutralizing actA antibody (anti-actA) in HH mice. c Renal actA is increased after UUO, with a greater induction in HH mice. Both are attenuated by actA neutralization. Boxed areas are shown at higher magnification immediately below. ActA increases are seen particularly in tubular epithelial cells. d Renal alpha SMA, fibronectin (FN), pSmad3 and MRTF-A are increased after UUO and this is augmented in HH kidneys. Expression of all is attenuated by actA neutralization in both WT and HH kidneys. (n = 6–9) *, **, ***, ****P < 0.05, 0.01, 0.001, 0.0001; one-way ANOVA with Tukey’s multiple comparisons post hoc test where there are > 2 groups; t-test for 2 groups Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36717814), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Western Blot

Detection of Activin A beta A subunit by Western Blot

Activin inhibition attenuates TGF beta 1-induced fibrotic responses and Smad3 activation in MC. Activin inhibition with follistatin (FST) decreases TGF beta 1-induced: a FN, alpha SMA and CTGF upregulation at 48 h (n = 5), b Smad3 phosphorylation (pSmad3) at 24 h (n = 5), c Smad3 nuclear translocation as assessed using eGFP-Smad3 (n = 3; 25–30 cells quantified per treatment group) at 24 h, and d Smad3 transcriptional activity at 24 h (n = 8). e Time course experiments show increases in pSmad3 occur earlier (30–60 min) with TGF beta 1 (n = 4) compared with actA (n = 4) or actB (n = 3) (18–48 h). *, **, ***, ****P < 0.05, 0.01, 0.001, 0.0001; one-way ANOVA with Tukey’s multiple comparisons post hoc test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36717814), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Western Blot

Detection of Activin A beta A subunit by Western Blot

Activin inhibition attenuates TGF beta 1-induced fibrotic responses and Smad3 activation in MC. Activin inhibition with follistatin (FST) decreases TGF beta 1-induced: a FN, alpha SMA and CTGF upregulation at 48 h (n = 5), b Smad3 phosphorylation (pSmad3) at 24 h (n = 5), c Smad3 nuclear translocation as assessed using eGFP-Smad3 (n = 3; 25–30 cells quantified per treatment group) at 24 h, and d Smad3 transcriptional activity at 24 h (n = 8). e Time course experiments show increases in pSmad3 occur earlier (30–60 min) with TGF beta 1 (n = 4) compared with actA (n = 4) or actB (n = 3) (18–48 h). *, **, ***, ****P < 0.05, 0.01, 0.001, 0.0001; one-way ANOVA with Tukey’s multiple comparisons post hoc test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36717814), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Activin A beta A subunit by Immunohistochemistry

Detection of Activin A beta A subunit by Immunohistochemistry

Activin A neutralization inhibits renal fibrosis in TGF beta 1-overexpressing mice. a TGF beta 1 transcript is increased in mice genetically engineered to overexpress TGF beta 1 (HH) compared with wild-type mice (WT) (n = 6–7, *p ≤ 0.05). b Serum actA is elevated in wild-type and HH mice after UUO. This is decreased by treatment with a neutralizing actA antibody (anti-actA) in HH mice. c Renal actA is increased after UUO, with a greater induction in HH mice. Both are attenuated by actA neutralization. Boxed areas are shown at higher magnification immediately below. ActA increases are seen particularly in tubular epithelial cells. d Renal alpha SMA, fibronectin (FN), pSmad3 and MRTF-A are increased after UUO and this is augmented in HH kidneys. Expression of all is attenuated by actA neutralization in both WT and HH kidneys. (n = 6–9) *, **, ***, ****P < 0.05, 0.01, 0.001, 0.0001; one-way ANOVA with Tukey’s multiple comparisons post hoc test where there are > 2 groups; t-test for 2 groups Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36717814), licensed under a CC-BY license. Not internally tested by R&D Systems.
Human Activin A ELISA Standard Curve

Human Activin A ELISA Standard Curve

Human Activin A was serially diluted and captured by Mouse Anti-Human/Mouse/Rat Activin A beta A subunit Monoclonal Antibody (Catalog # MAB3381) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human/Mouse/Rat Activin A beta A subunit Monoclonal Antibody (Catalog # MAB3381) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Activin A betaA subunit Antibody (69403)

Application
Recommended Usage

Neutralization

Measured by its ability to neutralize Activin A-induced hemoglobin expression in the K562 human chronic myelogenous leukemia cell line. The Neutralization Dose (ND50) is typically 0.02-0.06 µg/mL in the presence of 7.5 ng/mL Recombinant Human/Mouse/Rat Activin A.

Human/Mouse/Rat Activin A Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human/Mouse/Rat Activin A beta A subunit Biotinylated Antibody (Catalog # BAM3381)
  • Standard: Recombinant Human/Mouse/Rat Activin A Protein (Catalog # 338-AC)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using MAB3381 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Activin A betaA subunit

Activin and Inhibin are members of the TGF-beta superfamily of cytokines and are involved in a wide range of biological processes including tissue morphogenesis and repair, fibrosis, inflammation, neural development, hematopoiesis, reproductive system function, and carcinogenesis (1‑7). Activin and Inhibin are produced as precursor proteins. Their amino terminal propeptides are proteolytically cleaved and facilitate formation of disulfide-linked dimers of the bioactive proteins (8, 9). Activins are nonglycosylated homodimers or heterodimers of various beta  subunits ( beta A, beta B, beta C, and beta E in mammals), while Inhibins are heterodimers of a unique alpha  subunit and one of the beta  subunits. Activin A is a widely expressed homodimer of two beta A chains. The beta A subunit can also heterodimerize with a beta B or beta C subunit to form Activin AB and Activin AC, respectively (10). The 14 kDa mature human beta A chain shares 100% amino acid sequence identity with bovine, feline, mouse, porcine, and rat beta A. Activin A exerts its biological activities by binding to the type 2 serine/threonine kinase Activin RIIA which then noncovalently associates with the type 1 serine/threonine kinase Activin RIB/ALK-4 (7, 11). Signaling through this receptor complex leads to Smad activation and regulation of activin-responsive gene transcription (7, 11). The bioactivity of Activin A is regulated by a variety of mechanisms (11). BAMBI, Betaglycan, and Cripto are cell-associated molecules that function as decoy receptors or limit the ability of Activin A to induce receptor complex assembly (12‑14). The intracellular formation of Activin A can be prevented by the incorporation of the beta A subunit into Activin AC or Inhibin A (3, 10). And the bioavailability of Activin A is restricted by its incorporation into inactive complexes with alpha 2-Macroglobulin, Follistatin, and FLRG (15, 16).

References

  1. Kumanov, P. et al. (2005) Reprod. Biomed. Online 10:786.
  2. Maeshima, A. et al. (2008) Endocr. J. 55:1.
  3. Rodgarkia-Dara, C. et al. (2006) Mutat. Res. 613:123.
  4. Werner, S. and C. Alzheimer (2006) Cytokine Growth Factor Rev. 17:157.
  5. Xu, P. and A.K. Hall (2006) Dev. Biol. 299:303.
  6. Shav-Tal, Y. and D. Zipori (2002) Stem Cells 20:493.
  7. Chen, Y.G. et al. (2006) Exp. Biol. Med. 231:534.
  8. Gray, A.M. and A.J. Mason (1990) Science 247:1328.
  9. Mason, A.J. et al. (1996) Mol. Endocrinol. 10:1055.
  10. Thompson, T.B. et al. (2004) Mol. Cell. Endocrinol. 225:9.
  11. Harrison, C.A. et al. (2005) Trends Endocrinol. Metab. 16:73.
  12. Onichtchouk, D. et al. (1999) Nature 401:480.
  13. Gray, P.C. et al. (2002) Mol. Cell. Endocrinol. 188:254.
  14. Kelber, J.A. et al. (2008) J. Biol. Chem. 283:4490.
  15. Phillips, D.J. et al. (1997) J. Endocrinol. 155:65.
  16. Schneyer, A. et al. (2003) Endocrinology 144:1671.

Entrez Gene IDs

3624 (Human); 16323 (Mouse); 29200 (Rat)

Gene Symbol

INHBA

Additional Activin A betaA subunit Products

Product Documents for Activin A betaA subunit Antibody (69403)

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Product Specific Notices for Activin A betaA subunit Antibody (69403)

For research use only

Citations for Activin A betaA subunit Antibody (69403)

Customer Reviews for Activin A betaA subunit Antibody (69403) (2)

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  • Human/Mouse/Rat Activin A beta A subunit Antibody
    Name: Anonymous
    Application: Block/Neutralize
    Sample Tested: Serum-free Cell Culture Supernates
    Species: Human
    Verified Customer | Posted 07/07/2020
    We used this INHBA (100ng/mL) neutralizing antibody as part of an antibody cocktail containing several other antibodies to neutralize INHBA present in the Conditioned Medium (CM) from BT549 Cells treated with Angiotensin II. The effect of cytokines in the CM (+/-Neutralizing ab Cocktail) on Endothelial (HUVEC) Cell migration was measured using Incucyte Zoom Chemotaxis Assay.
    Activin A betaA subunit Antibody (69403) MAB3381
  • Human/Mouse/Rat Activin A beta A subunit Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Recombinant protein
    Species: Human
    Verified Customer | Posted 03/28/2019
    I added a sulfo tag to this antibody and used it as a detection antibody when developing an immunoassay for Activin A.
    Activin A betaA subunit Antibody (69403) MAB3381

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