Active Recombinant Human ACVR1, Fc-tagged, Biotinylated

• Cat.No.: ACVR1-526H
• Product Overview: The recombinant human ACVR1-Fc fusion protein is expressed as a 331amino acid protein consisting of Met21 - Glu123 region of ACVR1 (UniProt accession #Q04771) and a C-terminal Fc fusion from human IgG1, which exists as a dimer under non-reducing condition.

Specification

• Species: Human
• Source: Human Cells
• Tag: Fc
• Protein Length: 21-123 a.a.
• Form: Supplied at 0.5 mg/ml in sterile PBSpH7.4 (carrier & preservative free). The purified recombinant protein was labeled with Biotin (3-5 Biotin per molecule).
• Bio-activity: Immobilized ACVR1 protein binds human BMP6 in a functional ELISA. Blocks BMP6-mediated signaling activity.
• Molecular Mass: Calculated molecular mass (kDa): 37.1; Estimated by SDS-PAGE under reducing condition (kDa): ~45
• AA Sequence: MEDEKPKVNPKLYMCVCEGLSCGNEDHCEGQQCFSSLSINDGFHVYQKGCFQVYEQGKMTCKTPPSPGQAVECC QGDWCNRNITAQLPTKGKSFPGTQNFHLESTGTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
• Endotoxin: <0.1 eu per 1 μg of purified recombinant protein determined by the
• Purity: >95% judged by SDS-PAGE under reducing condition
• Storage: The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 3 months. The undiluted protein can be stored at 4°C for no more than 2 weeks. Avoid repeated freeze-thaw cycles.
• Publications:
  An anti-ACVR1 antibody exacerbates heterotopic ossification by fibro/adipogenic progenitors in fibrodysplasia ossificans progressiva mice (2021)

Gene Information

• Gene Name: ACVR1 activin A receptor, type I [ Homo sapiens ]
• Official Symbol: ACVR1
• Synonyms: ACVR1; activin A receptor, type I; ACVRLK2; activin receptor type-1; ACVR1A; ALK2; SKR1; activin receptor type I; hydroxyalkyl-protein kinase; activin receptor-like kinase 2; TGF-B superfamily receptor type I; activin A receptor, type II-like kinase 2; serine/threonine-protein kinase receptor R1; FOP; TSRI; ACTRI
• Gene ID: 90
• mRNA Refseq: NM_001105
• Protein Refseq: NP_001096
• MIM: 102576
• UniProt ID: Q04771
• Chromosome Location: 2q23-q24
• Pathway: ALK1 pathway, organism-specific biosystem; ALK1 signaling events, organism-specific biosystem; ALK2 signaling events, organism-specific biosystem; Cytokine-cytokine receptor interaction, organism-specific biosystem; Cytokine-cytokine receptor interaction, conserved biosystem; TGF-beta signaling pathway, organism-specific biosystem; TGF-beta signaling pathway, conserved biosystem
• Function: ATP binding; SMAD binding; activin binding; contributes_to activin receptor activity, type I; follistatin binding; metal ion binding; nucleotide binding; protein binding; protein homodimerization activity; protein serine/threonine kinase activity; receptor activity; receptor signaling protein serine/threonine kinase activity; transforming growth factor beta binding; transforming growth factor beta receptor activity, type I; transmembrane receptor protein serine/threonine kinase activity

Citation

An anti-ACVR1 antibody exacerbates heterotopic ossification by fibro-adipogenic progenitors in fibrodysplasia ossificans progressiva mice

Journal: The Journal of Clinical Investigation    PubMed ID: 35503416    Data: 2022/6/15

Authors: John B. Lees-Shepard, Sean J. Stoessel, David J. Goldhamer

Article Snippet:The phagemid was used to transform XL1-blue E . coli cells to generate the anti-ACVR1 library.The phagemid was used to transform XL1-blue E . coli cells to generate the anti-ACVR1 library.. The library was panned for 2 rounds on biotinylated ACVR1 ECD (Creative BioMart) that was conjugated to magnetic streptavidin beads (Dynabeads MyOne, Thermo Fisher Scientific, 65601).. Streptavidin beads without ACVR1 were used in deselection steps to remove nonspecific binders.Streptavidin beads without ACVR1 were used in deselection steps to remove nonspecific binders.

( A ) Mouse C2C12 myoblast cells express similar levels of Acvr1 and Alk3 , low levels of Alk1 , and no detectable Alk6 , as quantified by RT-qPCR ( n = 3). ΔCT values were calculated using the average CT values of the internal controls, Gapdh or Actb (β-actin) (see Methods). Error bars represent ±SD. CT values >40 were considered not detected (N.D.). ( B ) Surface expression of ACVR1 and ALK3 on C2C12 cells, as detected by flow cytometry. ( C ) mAb JAB0505 binds to parental C2C12 cells, but not Acvr1 -KO cells, as assessed by flow cytometry. ( D ) JAB0505 inhibits BMP9-induced signal activation in wild-type and ACVR1(R206H)-overexpressing C2C12 cells in a dose-dependent manner, as determined by quantification of BRE-luciferase activity ( n = 3). Error bars represent ±SD.

( A ) Representative μCT images of HO in Acvr1 FLEx(R206H)/+ ; CAG-Cre ERT2 mice 20 days after cardiotoxin-induced injury of the gastrocnemius muscle (Untreated, n = 3; JAB0505, n = 4). ( B ) Representative μCT images of HO (pseudocolored green) in Acvr1 tnR206H/+ ; Tie2-Cre mice 21 days after pinch injury of the gastrocnemius muscle (Untreated, n = 11; JAB0505, n = 10). ( C ) Quantification of HO volumes as a function of time after muscle pinch injury of Acvr1 tnR206H/+ ; Tie2-Cre mice. Untreated, n = 11; JAB0505 (10 mg/kg), n = 6. Error bars represent ±SEM. **** P ≤ 0.0001 by 2-way ANOVA with Sidak’s multiple-comparison test. ( D ) Paired single transverse slice and 3D reconstructed μCT images of the distal hind limb of Acvr1 tnR206H/+ ; Tie2-Cre mice at the indicated times after hind limb muscle pinch injury with and without administration of JAB0505. Mineralized bone in the slice images is pseudocolored green. Radio-opaque lesional tissue below the threshold set for quantification of mineralized bone (white arrows in day 14 slices) is extensive at day 14 in JAB0505-treated mice. Mineralized bone in day 14 slices is barely visible at this magnification. HO volumes are given for images prior to day 35. The tibia and fibula are labeled with asterisks in the day 14 slices. Pelvic bones present in day 21, 28, and 35 slices of JAB0505-treated mice are denoted with arrowheads. To avoid confusion with HO, the baculum present in some images was removed by segmentation.

( A ) Transverse sections of muscle from untreated and JAB0505-treated Acvr1 tnR206H/+ ; Tie2-Cre mice on days 6 and 14 after muscle pinch injury. Alcian blue staining to detect cartilage (blue) and immunohistochemical staining to detect ACVR1 (brown) were performed on nearby sections. Sections processed for Alcian blue were counterstained with eosin, and sections processed for ACVR1 immunohistochemistry were counterstained with hematoxylin. On day 6 after injury, untreated Acvr1 tnR206H/+ ; Tie2-Cre mice exhibited a spatially discrete lesional region (asterisk) that was primarily comprised of ACVR1-positive ectopic cartilage. By day 14, the lesional region (asterisk) of untreated Acvr1 tnR206H/+ ; Tie2-Cre mice displayed sporadic ACVR1 localization and was composed of both cartilage and morphologically apparent bone. In contrast, JAB0505 treated Acvr1 tnR206H/+ ; Tie2-Cre mice displayed multiple apparent cartilaginous lesions and broader distribution of ACVR1 localization on days 6 and 14 (arrows). Centrally located myofiber nuclei (arrowheads), which identify regenerated fibers, were rare in Acvr1 tnR206H/+ ; Tie2-Cre mice, and undetected in JAB0505-treated Acvr1 tnR206H/+ ; Tie2-Cre mice. AB/E, Alcian blue/eosin. Original magnification, ×100. ( B ) Transverse sections of lower hind limbs of Acvr1 tnR206H/+ ; Tie2-Cre mice 14 days after injury. Alcian blue staining revealed numerous cartilaginous lesions (blue, examples at arrows) in injured muscle of JAB0505-treated mice. Sections were counterstained with eosin. T, tibia. Original magnification, ×40.