Human/Mouse/Rat GDF-8/Myostatin Antibody AF788: R&D Systems

Human/Mouse/Rat GDF-8/Myostatin Antibody

(4 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat GDF-8/Myostatin in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 30% cross‑reactivity is observed with recombinant human/mouse/rat GDF-11. In Western blots, no cross-reactivity with recombinant human BMP‑6 and recombinant mouse BMP-6 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant mouse GDF-8/Myostatin
    Asp268-Ser376
    Accession # O08689
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize GDF‑8/Myostatin-induced hemoglobin expression in the K562 human chronic myelogenous leukemia cell line. The Neutralization Dose (ND50) is typically 0.6-3 µg/mL in the presence of 30 ng/mL Recombinant Mouse GDF‑8/Myostatin.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Recombinant Human, Mouse, and Rat GDF‑8/Myostatin by Western Blot. Western blot shows 25 ng of Recombinant Human/Mouse/Rat GDF‑8/Myostatin (Catalog # 788‑G8), Recombinant Human/Mouse/Rat GDF‑11/BMP‑11 (Catalog # 1958‑GD), Recombinant Human BMP‑6 (Catalog # 507‑BP), and Recombinant Mouse BMP‑6 (Catalog # 6325‑BM). PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human/Mouse/Rat GDF‑8/Myostatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF788) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for GDF‑8/Myostatin at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Immunohistochemistry
GDF‑8/Myostatin in Mouse Embryo. GDF‑8/Myostatin was detected in immersion fixed frozen sections of mouse embryo (10 d.p.c., section through neural tube) using Goat Anti-Human/Mouse/Rat GDF‑8/Myostatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF788) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Hemoglobin Expression Induced by GDF‑8/Myostatin and Neutralization by Mouse GDF‑8/Myostatin Antibody. Recombinant Mouse
GDF‑8/Myostatin (Catalog # 788-G8) 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 Mouse GDF‑8/Myostatin (30 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Mouse/Rat GDF‑8/Myostatin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF788). The ND50 is typically 0.6-3 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
Background: GDF-8/Myostatin
Growth Differentiation Factor 8 (GDF-8), also known as myostatin, is a member of the TGF-beta superfamily that is expressed specifically in developing and adult skeletal muscle. GDF-8 cDNA encodes a 376 amino acid (aa) prepropeptide with a 24 aa residue signal peptide, a 223 aa residue amino-terminal propeptide, and a 109 aa residue carboxy-terminal mature protein. Mature GDF-8 contains the canonical 7-cysteine motif common to other TGF-beta superfamily members. Similar to the TGF-beta s, activins and BMP-11, GDF-8 also contains one extra pair of cysteine residues that is not found in other family members. The bioactive form of GDF-8 is a homodimer with an apparent molecular weight of approximately 25 kDa. GDF-8 is highly conserved across species. At the amino acid sequence level, mature human, mouse, rat and cow GDF-8 are 100% identical. Within the TGF-beta superfamily, GDF-8 is most closely related to BMP-11, a mammalian protein that acts as a dorsal mesoderm and neural inducer in Xenopus explants. The two proteins share 90% amino acid sequence identity within their mature chain. A targeted disruption of GDF-8 in mouse results in large mice with a widespread increase in skeletal muscle mass, indicating that GDF-8 is a negative regulator of skeletal muscle growth. A mutation in the bovine GDF-8 gene has been shown to be responsible for the double-muscled phenotype in cattle breeds such as Belgian Blue cattle that is characterized by an increase in muscle mass. GDF-8 has also been shown to inhibit preadipocyte differentiation to adipocytes. Mature GDF-8 binds to activin type II receptors and the binding is antagonized by the activin-binding protein, follistatin. R&D Systems recombinant GDF-8 preparations have been shown to act similarly to Activin A in both the Xenopus animal cap and the K562 assays.
  • References:
    1. Storm, E.E. et al. (1994) Nature 368:639.
    2. Sharma, M. et al. (1999) J. Cell Physiol. 180:1.
    3. McPherron, A.C. et al. (1997) Nature 387:83.
    4. Lee, S.J. et al. (2001) Proc. Natl. Acad. Sci. USA 98:9306.
    5. Kim, H.S. et al. (2001) Biochem. Biophys. Res. Commun. 281:902.
  • Long Name:
    Growth Differentiation Factor 8
  • Entrez Gene IDs:
    2660 (Human); 17700 (Mouse)
  • Alternate Names:
    GDF8; GDF-8; GDF8growth differentiation factor 8; growth/differentiation factor 8; MSTN; Myostatin
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Compensatory anabolic signaling in the sarcopenia of experimental chronic arthritis
    Authors: RD Little, I Prieto-Pot, S Pérez-Baos, A Villalvill, P Gratal, F Cicuttini, R Largo, G Herrero-Be
    Sci Rep, 2017;7(1):6311.
    Species: Rabbit
    Sample Type: Tissue Homogenates
    Application: WB
  2. A single heterochronic blood exchange reveals rapid inhibition of multiple tissues by old blood
    Nat Commun, 2016;7(0):13363.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Fresh
  3. High concentrations of HGF inhibit skeletal muscle satellite cell proliferation in vitro by inducing expression of myostatin: a possible mechanism for reestablishing satellite cell quiescence in vivo.
    Authors: Yamada M, Tatsumi R, Yamanouchi K, Hosoyama T, Shiratsuchi S, Sato A, Mizunoya W, Ikeuchi Y, Furuse M, Allen RE
    Am. J. Physiol., Cell Physiol., 2010;298(3):C465-76.
    Species: Rat
    Sample Type: Whole Cells
    Application: ICC
  4. Increased secretion and expression of myostatin in skeletal muscle from extremely obese women.
    Authors: Hittel DS, Berggren JR, Shearer J, Boyle K, Houmard JA
    Diabetes, 2009;58(1):30-8.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: WB
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