Human/Mouse/Rat GDF-8/Myostatin Antibody Summary
Asp268-Ser376
Accession # O08689
Applications
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.
Scientific Data
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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 (788-G8), Recombinant Human/Mouse/Rat GDF-11/BMP-11 (Catalog # 1958-GD), Recombinant Human BMP-6 (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.
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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; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
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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.
Reconstitution Calculator
Preparation and Storage
- 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.
- Storm, E.E. et al. (1994) Nature 368:639.
- Sharma, M. et al. (1999) J. Cell Physiol. 180:1.
- McPherron, A.C. et al. (1997) Nature 387:83.
- Lee, S.J. et al. (2001) Proc. Natl. Acad. Sci. USA 98:9306.
- Kim, H.S. et al. (2001) Biochem. Biophys. Res. Commun. 281:902.
Product Datasheets
Citations for Human/Mouse/Rat GDF-8/Myostatin Antibody
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.
23
Citations: Showing 1 - 10
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The relationship between myodural bridge, atrophy and hyperplasia of the suboccipital musculature, and cerebrospinal fluid dynamics
Authors: Yang, H;Wei, XS;Gong, J;Du, XM;Feng, HB;Su, C;Gilmore, C;Yue, C;Yu, SB;Li, C;Sui, HJ;
Scientific reports
Species: Rat
Sample Types: Tissue Homogenates
Applications: Western Blot -
Effects of Tofacitinib on Muscle Remodeling in Experimental Rheumatoid Sarcopenia
Authors: Bermejo-Álvarez, I;Pérez-Baos, S;Gratal, P;Medina, JP;Largo, R;Herrero-Beaumont, G;Mediero, A;
International journal of molecular sciences
Species: Rabbit
Sample Types: Tissue Homogenates
Applications: Western Blot -
Reversible Effects of Functional Mandibular Lateral Shift on Masticatory Muscles in Growing Rats
Authors: Guan, H;Yonemitsu, I;Ikeda, Y;Ono, T;
Biomedicines
Species: Rat
Sample Types: Whole Tissue
Applications: IHC -
Myostain is involved in ginsenoside Rb1-mediated anti-obesity
Authors: Hong-Shi Li, Jiang-Ying Kuang, Gui-Jun Liu, Wei-Jie Wu, Xian-Lun Yin, Hao-Dong Li et al.
Pharmaceutical Biology
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Maternal rodent exposure to di‐(2‐ethylhexyl) phthalate decreases muscle mass in the offspring by increasing myostatin
Authors: Fengju Li, Ting Luo, Honghui Rong, Lu Lu, Ling Zhang, Chuanfeng Zheng et al.
Journal of Cachexia, Sarcopenia and Muscle
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The relationship between myodural bridges, hyperplasia of the suboccipital musculature, and intracranial pressure
Authors: C Li, C Yue, ZC Liu, J Gong, XS Wei, H Yang, C Gilmore, SB Yu, GD Hack, HJ Sui
PLoS ONE, 2022-09-02;17(9):e0273193.
Species: Rat
Sample Types: Tissue Homogenates
Applications: Western Blot -
Novel myostatin-specific antibody enhances muscle strength in muscle disease models
Authors: H Muramatsu, T Kuramochi, H Katada, A Ueyama, Y Ruike, K Ohmine, M Shida-Kawa, R Miyano-Nis, Y Shimizu, M Okuda, Y Hori, M Hayashi, K Haraya, N Ban, T Nonaka, M Honda, H Kitamura, K Hattori, T Kitazawa, T Igawa, Y Kawabe, J Nezu
Scientific Reports, 2021-01-25;11(1):2160.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Western Blot -
A GDF11/myostatin inhibitor, GDF11 propeptide-Fc, increases skeletal muscle mass and improves muscle strength in dystrophic mdx mice
Authors: Q Jin, C Qiao, J Li, B Xiao, J Li, X Xiao
Skelet Muscle, 2019-05-27;9(1):16.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Impact of heat therapy on recovery after eccentric exercise in humans
Authors: Kyoungrae Kim, Shihuan Kuang, Qifan Song, Timothy P. Gavin, Bruno T. Roseguini
Journal of Applied Physiology
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Brown Adipose Tissue Controls Skeletal Muscle Function via the Secretion of Myostatin
Authors: Xingxing Kong, Ting Yao, Peng Zhou, Lawrence Kazak, Danielle Tenen, Anna Lyubetskaya et al.
Cell Metabolism
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Twist1 Activation in Muscle Progenitor Cells Causes Muscle Loss Akin to Cancer Cachexia
Authors: Parash Parajuli, Santosh Kumar, Audrey Loumaye, Purba Singh, Sailaja Eragamreddy, Thien Ly Nguyen et al.
Developmental Cell
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Inhibition of GDF8 (Myostatin) accelerates bone regeneration in diabetes mellitus type 2
Authors: C Wallner, H Jaurich, JM Wagner, M Becerikli, K Harati, M Dadras, M Lehnhardt, B Behr
Sci Rep, 2017-08-29;7(1):9878.
Species: Mouse
Sample Types: Whole Cells
Applications: Neutralization -
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, 2017-07-24;7(1):6311.
Species: Rabbit
Sample Types: Tissue Homogenates
Applications: Western Blot -
Myostatin signaling is up-regulated in female patients with advanced heart failure
Authors: J Ishida, M Konishi, M Saitoh, M Anker, SD Anker, J Springer
Int. J. Cardiol., 2017-04-04;0(0):.
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot -
The Use of Platelet-Rich and Platelet-Poor Plasma to Enhance Differentiation of Skeletal Myoblasts: Implications for the Use of Autologous Blood Products for Muscle Regeneration
Authors: O Miroshnych, WT Chang, JL Dragoo
Am J Sports Med, 2016-12-27;45(4):945-953.
Species: Human
Sample Types: Protein
Applications: Western Blot -
A single heterochronic blood exchange reveals rapid inhibition of multiple tissues by old blood
Nat Commun, 2016-11-22;7(0):13363.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-Fr -
Catabolic signaling and muscle wasting after acute ischemic stroke in mice: indication for a stroke-specific sarcopenia
Authors: Jochen Springer, Susanne Schust, Katrin Peske, Anika Tschirner, Andre Rex, Odilo Engel et al.
Stroke
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Latent myostatin has significant activity and this activity is controlled more efficiently by WFIKKN 1 than by WFIKKN 2
Authors: György Szláma, Mária Trexler, László Patthy
The FEBS Journal
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Immunolocalization of Myostatin (GDF-8) Following Musculoskeletal Injury and the Effects of Exogenous Myostatin on Muscle and Bone Healing
Authors: Moataz Elkasrawy, David Immel, Xuejun Wen, Xiaoyan Liu, Li-Fang Liang, Mark W. Hamrick
Journal of Histochemistry & Cytochemistry
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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., 2009-12-09;298(3):C465-76.
Species: Rat
Sample Types: Whole Cells
Applications: ICC -
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, 2008-10-03;58(1):30-8.
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot -
MT-102 prevents tissue wasting and improves survival in a rat model of severe cancer cachexia
Authors: POtsch MS, Ishida J, Palus S et al.
J Cachexia Sarcopenia Muscle
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The anabolic catabolic transforming agent (ACTA) espindolol increases muscle mass and decreases fat mass in old rats.
Authors: Potsch MS, Tschirner A, Palus S et al.
J Cachexia Sarcopenia Muscle.
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The test samples consisted of skeletal muscle tissue homogenates derived from mouse and avian muscle tissues. The homogenisation buffer contained Roche mini cOmplete protease inhibitor cocktail. The positive control used was 1μg of R&D systems recombinant myostatin/GDF8 (#788-G8).
I had loaded 20μg of total protein for each of the avian and mouse samples, along with1μg of R&D systems recombinant myostatin/GDF8 (#788-G8) into a Bio-Rad PROTEAN TGX 4-15% gel.
The electrophoresis was performed with NuPAGE MES running buffer at 90V constant current, and the proteins were subsequently blotted onto a 0.2μm nitrocellulose membrane. To check for transfer efficiency and protein integrity, i had stained the blot with Ponceau S and found that the transfer was efficient and the proteins were not degraded. Subsequently, i destained the blot with TBS-T and deionised water, and blocked the blots with 5% milk in TBS-T.
Prior to incubating the membrane in primary antibody, i had cut the membrane such that one section contained all the animal tissue samples and the other section contained the recombinant GDF-8/myostatin and the MagicMark XP western standard.
The cut sections were incubated separately with identical concentrations of primary and secondary antibodies - R&D systems goat polyclonal GDF8 antibody (AF788-SP) at 2 µg/mL concentration, and R&D system anti-goat secondary antibody (HAF017) at 1:4000 dilution.
The recombinant GDF-8/Myostatin positive control and the MagicMark XP western standards produced clear and robust signals.
However, the sections containing the samples derived from animal tissues (avian and mouse) failed to produce any discernible signal.
I had repeated the same experiment above three times with increasing concentrations of primary antibody - 0.1, 0.25 and 2 µg/mL. However, all three experiments yielded the same result - strong signal for the recombinant GDF-8/Myostatin, but no visible signal detected with the avian and mouse tissue samples.
