Rat 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 9q22. 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 55 53 50 52 52 1.80 3.4% Sample 2 228 261 228 264 245 17.28 7.0% Sample 3 845 788 838 845 829 23.84 2.8% 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 55 53 50 52 52 1.80 3.4% Sample 1 55 53 50 52 52 1.80 3.4% Sample 2 228 261 228 264 245 17.28 7.0% Sample 2 228 261 228 264 245 17.28 7.0% Sample 3 845 788 838 845 829 23.84 2.8% Sample 3 845 788 838 845 829 23.84 2.8%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 55 228 845 60 239 884 Standard deviation 2.31 14.36 38.03 3.18 15.30 63.65 CV (%) 4.2% 6.3% 4.5% 5.3% 6.4% 7.2% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 55 228 845 60 239 884 Standard deviation 2.31 14.36 38.03 3.18 15.30 63.65 CV (%) 4.2% 6.3% 4.5% 5.3% 6.4% 7.2% 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) 55 228 845 60 239 884 Mean (pg/ml) 55 228 845 60 239 884 Standard deviation 2.31 14.36 38.03 3.18 15.30 63.65 Standard deviation 2.31 14.36 38.03 3.18 15.30 63.65 CV (%) 4.2% 6.3% 4.5% 5.3% 6.4% 7.2% CV (%) 4.2% 6.3% 4.5% 5.3% 6.4% 7.2%

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. 0.047 0.133 0.21 0.322 0.53 0.885 1.511 2.395 Concentration (pg/ml) 0 31.2 62.5 125 250 500 1000 2000 O.D. 0.047 0.133 0.21 0.322 0.53 0.885 1.511 2.395 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. 0.047 0.133 0.21 0.322 0.53 0.885 1.511 2.395 O.D. 0.047 0.133 0.21 0.322 0.53 0.885 1.511 2.395

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.