polyclonal rabbit anti human myom3 antibody  (Proteintech)

Journal: Wellcome Open Research

Article Title: Evaluation of a six-minute walk test in the DE50-MD canine model of Duchenne muscular dystrophy and its effect on blood-borne biomarkers

doi: 10.12688/wellcomeopenres.23269.2

Figure Lengend Snippet: Samples analysed for each of the assays performed on blood samples collected before and after a 6MWT in 12 month old dogs. Dog ID abbreviations indicate the genotype and the individual dog code: abbreviations beginning with WT are wild-type dogs and those beginning with DE50 are dystrophic DE50-MD dogs. Analytes measured were CK activity in plasma, and myomesin 3 (MYOM3), myostatin (MSTN), Luminex cytokine/chemokine panel, and dystromiRs (miR-1, miR-133a and miR-206) in serum. Boxes shaded in grey represent samples not available for analysis.

Article Snippet: Membranes were incubated with a polyclonal rabbit anti-human MYOM3 antibody (Proteintech, #17692-1-AP, 1:1000 dilution) and a polyclonal rabbit anti-canine albumin antibody (as internal loading control, Biorbyt Ltd, #orb242465; 1:1,000,000 dilution) overnight at 4°C.

Techniques: Activity Assay, Clinical Proteomics, Luminex

Journal: Wellcome Open Research

Article Title: Evaluation of a six-minute walk test in the DE50-MD canine model of Duchenne muscular dystrophy and its effect on blood-borne biomarkers

doi: 10.12688/wellcomeopenres.23269.2

Figure Lengend Snippet: Results for analytes of interest for WT (blue) and DE50-MD dogs (red). A ) CCL2 Luminex assay results (ng/ml), B ) MYOM3 western blot results normalised to albumin (AU), C ) IL-10 Luminex assay results (ng/ml), D ) miR-1 RT-qPCR log relative quantity, E ) miR-133a RT-qPCR log relative quantity, F ) miR-206 RT-qPCR log relative quantity, G ) MSTN ELISA results (pg/ml), H ) CK activity (Units/L), I ) KC-LIKE Luminex assay results (ng/ml), J ) percentage change in CK activity 3-hours post-6MWT, K ) percentage change in KC-LIKE 3-hours post-6MWT. DE50-MD N=8, WT N=4 for graphs A – C and G – K ; DE50-MD N=6, WT N=2 for graphs D – F . Boxes extend from the 25 th to 75 th percentile, with a line within the box at the median value. Each point represents an individual DE50-MD or WT dog, and whiskers show the minimum and maximum results for that age-group. Letters a, b and c denote statistically significant differences (P<0.05) in the mean within the DE50-MD (red letters) or WT (blue letters) genotype: means sharing a letter are not significantly different. Asterisks denote the level of significance of a difference between genotypes based on linear mixed model analysis, adjusted for repeated measures: * P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: Membranes were incubated with a polyclonal rabbit anti-human MYOM3 antibody (Proteintech, #17692-1-AP, 1:1000 dilution) and a polyclonal rabbit anti-canine albumin antibody (as internal loading control, Biorbyt Ltd, #orb242465; 1:1,000,000 dilution) overnight at 4°C.

Techniques: Luminex, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Activity Assay

Journal: Wellcome Open Research

Article Title: Longitudinal assessment of blood-borne musculoskeletal disease biomarkers in the DE50-MD dog model of Duchenne muscular dystrophy

doi: 10.12688/wellcomeopenres.17398.2

Figure Lengend Snippet: Figure a ) representative MYOM3 western blot. This western blot shows a positive control mini-dilution series that was run alongside samples in every gel, followed by 5 samples of WT control dog serum, 5 samples of DE50-MD dog serum, and a negative control lane in which no protein was loaded. Figure b ) MYOM3 quantified by western blot in serum samples from dogs studied longitudinally between 1 and 18 months of age. DE50-MD: grey, total N=83 samples, from N=10 dogs, N=2-8 per age group; WT dogs: (not shown, see Underlying data ), total N=113 samples, from N=10 dogs, N=3-9 per age group. Bands corresponding to MYOM3 were not detected in any WT serum samples tested. Boxes extend from the 25 th to 75 th percentile, with a line within the box at the median value. Each point represents an individual sample, and whiskers show the minimum and maximum results for that age-group. Band intensity calculated by densitometry using ImageJ software. Figure c ) correlation between MYOM3 quantity and CK activity in blood samples from DE50-MD dogs studied longitudinally between 1 and 18 months, total of N=75 samples, from N=10 dogs. (MYOM3: myomesin-3; WT: wild type.)

Article Snippet: Membranes were blocked in 10% milk powder (Marvel) in phosphate buffered saline (PBS, Fisher Scientific, #BR0014G)/Tween 0.05% (Sigma, #P9416) for one hour at room temperature, before incubation with a polyclonal rabbit anti-human MYOM3 antibody (Proteintech, #17692-1-AP, 1:1000 dilution) and a polyclonal rabbit anti-canine albumin antibody (as internal loading control, Biorbyt Ltd, #orb242465; 1:1,000,000 dilution) overnight at 4 °C.

Techniques: Western Blot, Positive Control, Control, Negative Control, Software, Activity Assay

Journal: Wellcome Open Research

Article Title: Longitudinal assessment of blood-borne musculoskeletal disease biomarkers in the DE50-MD dog model of Duchenne muscular dystrophy

doi: 10.12688/wellcomeopenres.17398.2

Figure Lengend Snippet: Included in the analysis was plasma CK activity (log10 U/l), serum MYOM3 quantity (AU), serum MSTN concentration (ng/ml), serum miR-1, miR-133a and miR 206 levels (log10 relative quantity). Study population consisted of DE50-MD and wild-type (WT) dogs studied longitudinally between 3 and 18 months of age. DE50-MD: grey, N=10 dogs total, N=2-9 per age group; WT: white, N=7 dogs total, N=2-5 per age group. Where an individual was missing data for a specific biomarker, missing data was replaced with the mean of the relevant genotype and age-group for that biomarker. Boxes extend from the 25 th to 75 th percentile, with a line within the box at the median value. Each point represents an individual sample, and whiskers show the minimum and maximum results for that age-group. Asterisks denote the level of significance of a difference between genotypes based on linear mixed model analysis: ****P<0.0001. Letters a, b and c denote statistically significant differences (P<0.05) in the mean within either the DE50-MD (grey letters) or WT (black letters) genotypes: means sharing a letter are not significantly different within each genotype group. (CK: creatine kinase; WT: wild type.)

Article Snippet: Membranes were blocked in 10% milk powder (Marvel) in phosphate buffered saline (PBS, Fisher Scientific, #BR0014G)/Tween 0.05% (Sigma, #P9416) for one hour at room temperature, before incubation with a polyclonal rabbit anti-human MYOM3 antibody (Proteintech, #17692-1-AP, 1:1000 dilution) and a polyclonal rabbit anti-canine albumin antibody (as internal loading control, Biorbyt Ltd, #orb242465; 1:1,000,000 dilution) overnight at 4 °C.

Techniques: Clinical Proteomics, Activity Assay, Concentration Assay, Biomarker Discovery

Journal: Wellcome Open Research

Article Title: Longitudinal assessment of blood-borne musculoskeletal disease biomarkers in the DE50-MD dog model of Duchenne muscular dystrophy

doi: 10.12688/wellcomeopenres.17398.2

Figure Lengend Snippet: Sample size calculations. N required to show with sufficient power (0.8) an improvement in a DE50-MD biomarker towards WT levels with any given treatment. Sample sizes were calculated for the principal component analysis output (performed on blood-borne CK activity, MYOM3 quantity, MSTN concentration, and miR-1, -133a and -206 relative quantity) and for each individual biomarker that was significantly elevated in DE50-MD compared to WT blood samples for dogs aged 3–18 months. (CK: creatine kinase; MSTN: myostatin; MYOM3: myomesin-3; WT: wild type.)

Article Snippet: Membranes were blocked in 10% milk powder (Marvel) in phosphate buffered saline (PBS, Fisher Scientific, #BR0014G)/Tween 0.05% (Sigma, #P9416) for one hour at room temperature, before incubation with a polyclonal rabbit anti-human MYOM3 antibody (Proteintech, #17692-1-AP, 1:1000 dilution) and a polyclonal rabbit anti-canine albumin antibody (as internal loading control, Biorbyt Ltd, #orb242465; 1:1,000,000 dilution) overnight at 4 °C.

Techniques: Biomarker Discovery, Activity Assay, Concentration Assay