Human Myostatin/GDF8 ELISA Kit PicoKine®

Myostatin (also known as growth differentiation factor 8, abbreviated GDF-8) is a myokine, a protein produced and released by myocytes that acts on muscle cells' autocrine function to inhibit myogenesis: muscle cell growth and differentiation. In humans it is encoded by the MSTN gene. It is mapped to 2q32.2. Myostatin is a secreted growth differentiation factor that is a member of the TGF beta protein family. This gene encodes a secreted ligand of the TGF-beta (transforming growth factor-beta) superfamily of proteins. Ligands of this family bind various TGF-beta receptors leading to recruitment and activation of SMAD family transcription factors that regulate gene expression. The encoded preproprotein is proteolytically processed to generate each subunit of the disulfide-linked homodimer. This protein negatively regulates skeletal muscle cell proliferation and differentiation. Mutations in this gene are associated with increased skeletal muscle mass in humans and other mammals.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 51 51 50 50 50 0.50 1.0% Sample 2 171 174 183 172 175 4.74 2.7% Sample 3 628 577 554 606 591 28.10 4.7% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 51 51 50 50 50 0.50 1.0% Sample 1 51 51 50 50 50 0.50 1.0% Sample 2 171 174 183 172 175 4.74 2.7% Sample 2 171 174 183 172 175 4.74 2.7% Sample 3 628 577 554 606 591 28.10 4.7% Sample 3 628 577 554 606 591 28.10 4.7%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 51 171 628 61 181 638 Standard deviation 2.14 10.43 28.26 3.29 13.21 42.11 CV (%) 4.2% 6.1% 4.5% 5.4% 7.3% 6.6% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 51 171 628 61 181 638 Standard deviation 2.14 10.43 28.26 3.29 13.21 42.11 CV (%) 4.2% 6.1% 4.5% 5.4% 7.3% 6.6% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 51 171 628 61 181 638 Mean (pg/ml) 51 171 628 61 181 638 Standard deviation 2.14 10.43 28.26 3.29 13.21 42.11 Standard deviation 2.14 10.43 28.26 3.29 13.21 42.11 CV (%) 4.2% 6.1% 4.5% 5.4% 7.3% 6.6% CV (%) 4.2% 6.1% 4.5% 5.4% 7.3% 6.6%

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.07 0.196 0.278 0.413 0.627 0.973 1.633 2.359 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.07 0.196 0.278 0.413 0.627 0.973 1.633 2.359 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.07 0.196 0.278 0.413 0.627 0.973 1.633 2.359 O.D. 0.07 0.196 0.278 0.413 0.627 0.973 1.633 2.359

Activate the sample Cell culture supernatants: add activating reagent pro rata, i.e. add 50μl of Solution A into 100μl of sample, 10 min later, add 50μl of Solution B. PH 7.0-7.6. Serum, plasma (heparin, EDTA) : add activating reagent pro rata, i.e. add 20μl of Solution A into 40μl of sample, 10 min later, add 20μl of Solution B. PH 7.0-7.6. It is unnecessary to activate the recombinant Myostatin/GDF8. Sample was diluted partly after adding activating reagent, so please pay attention to this when calculate target protein concentration. Activate the sample Cell culture supernatants: add activating reagent pro rata, i.e. add 50μl of Solution A into 100μl of sample, 10 min later, add 50μl of Solution B. PH 7.0-7.6. Serum, plasma (heparin, EDTA) : add activating reagent pro rata, i.e. add 20μl of Solution A into 40μl of sample, 10 min later, add 20μl of Solution B. PH 7.0-7.6. It is unnecessary to activate the recombinant Myostatin/GDF8. Sample was diluted partly after adding activating reagent, so please pay attention to this when calculate target protein concentration.

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.