rabbit polyclonal anti myom3 antibody  (Proteintech)

Journal: Molecular Therapy. Nucleic Acids

Article Title: Improving angiogenesis ameliorates the efficacy of ASO-based exon skipping for the treatment of Duchenne muscular dystrophy

doi: 10.1016/j.omtn.2026.102834

Figure Lengend Snippet: Combined treatment mLVRF+ASO improves dystrophin restoration (A) Quantification of ASO in TA (tibialis anterior), GAS (gastrocnemius), QUAD (quadriceps), TRI (triceps), and DIA (diaphragm) after 12 weeks of ASO treatment (∗∗∗ p < 0.001 analyzed by two-way ANOVA). (B) Quantification by qPCR of exon 23 skipping in different muscles (∗ p < 0.05 analyzed by two-way ANOVA). (C) Representative WB from diaphragm protein lysates. A standard curve of 0%, 5%, 15%, and 20% was made from pooled lysates from C57Bl10 (WT) and mdx for each tissue. (D) Quantification of dystrophin expression by WB (∗ p < 0.05 analyzed by two-way ANOVA). (E) Dystrophin immunostaining on cross-sections of muscles from WT and scramble-, mLVRF-, ASO-, and mLVRF+ASO-treated mice. Scale bars, 100 μm. (F) Proportion of dystrophin-positive fibers in GAS muscles (∗∗ p < 0.01, analyzed by t test). (G) Western blot quantification of myomesin-3 (MYOM3) in serum after normalization to scramble group. Myomesin 3 is undetectable in the serum of WT mice. (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; analyzed by one-way ANOVA). Results are expressed as the mean ± SEM ( n = 4–5 per group).

Article Snippet: Membranes were incubated with a rabbit polyclonal anti-MYOM3 antibody (1:1,000, Proteintech, Rosemont, USA), followed by IRDye 800CW goat anti-rabbit IgG secondary antibody (1:2,000, LI-COR, Lincoln, USA).

Techniques: Muscles, Expressing, Immunostaining, Western Blot

Journal: Science Advances

Article Title: Lysosomal damage is a therapeutic target in Duchenne muscular dystrophy

doi: 10.1126/sciadv.adv6805

Figure Lengend Snippet: ( A ) Schematic representation of the study setup. Six-week-old Dmd mdx mice were injected intravenously with a rAAV encapsulating a μDys encoding sequence, under the control of an spc512 promoter, at two doses of 5 × 10 12 vg/kg (low dose) and 1 × 10 13 vg/kg (high dose). Four-week-old Sgcg −/− mice were injected intravenously with a rAAV9 encapsulating the human sequence of SGCG, under the control of a desmin promoter, at a dose of 2 × 10 13 vg/kg. IV, intravenous. ( B ) Dosage of CK in serum of mice at the end of the study (average ± SD). ( C ) ELISA quantification of MYOM3 in the serum of the mice before euthanasia. Data are presented as the scatterplot (average ± SD). ( D ) Global force evaluation by an escape test (force in newton normalized to body weight in grams) before euthanasia of mice (average ± SD). ( E ) Relative Lgals3 mRNA expression in the GA normalized to Rplp0 (average ± SD). ( F ) Representative confocal images of transversal sections of GA muscle immunostained for LAMP2 and LGALS3. Scale bars, 25 μm. ( G ) Quantification of double-positive puncta (LAMP2 + LGALS3 + ) in GA myofibers (median and range shown). An ANOVA test with Tukey’s multiple comparisons test was used for statistical comparisons. * P < 0.05, ** P < 0.05, *** P < 0.001, and **** P < 0.0001. Gray asterisks represent relative comparison to the “WT_Phy Ser” group, and red asterisks represent comparison to the “mdx_Phy Ser” group.

Article Snippet: An anti-MYOM3 polyclonal capture antibody (Proteintech Lab, ref. 17692-1-AP) was coated to the wells of a multiarray plate containing electrodes (MSD, ref. L15XA-3) (overnight, 4°C).

Techniques: Injection, Sequencing, Control, Enzyme-linked Immunosorbent Assay, Expressing, Comparison

Journal: Science Advances

Article Title: Lysosomal damage is a therapeutic target in Duchenne muscular dystrophy

doi: 10.1126/sciadv.adv6805

Figure Lengend Snippet: ( A ) Study setup. Dmd mdx-4Cv mice were started on TR treatment (2% dilution in drinking water) at 3 weeks. For groups 4 and 5, mice were injected intravenously with a rAAV9 encapsulating the μDys sequence at a dose of 7 × 10 12 vg/kg. ( B ) Seric CK analysis before the escape test (average ± SD, n = 5). ( C ) ELISA quantification of MYOM3 before the escape test (average ± SD, n = 5). ( D ) Global force evaluation (normalized to mice body weight, N/g) by the escape test (average ± SD, n = 5). ( E ) Histological characterization of muscles. Top panel: H&E labeling of GA muscle cross sections. Middle panel: Sirius red labeling of diaphragm and GA muscles. Bottom panel: Representative images of GA muscle serial cross sections immunostained for mouse immunoglobulin (IgG), laminin, and CD11b. Scale bars, 200 μm (H&E) and 500 and 100 μm (zoomed-in images). ( F ) Analysis of myofiber size distribution in GA muscles represented by the variance coefficient calculated as [SD of the muscle fiber size/mean of muscle fiber size] * 1000 (median with min-max values). ( G ) Centronucleation index in the GA muscle (median with min-max values). ( H and I ) Fibrosis analysis of GA and diaphragm by the quantification of Sirius red–positive areas on transversal sections (average ± SD). ( J ) Quantification of IgG uptake by myofibers (positive myofiber number normalized to muscle cross-sectional area). ( K ) Evaluation of CD11b + cell infiltration in the muscle by the quantification of CD11b area. An ANOVA test with Tukey’s multiple comparisons test was used for statistical comparisons (compared to WT control and mdx control). ANOVA: * P < 0.05, ** P < 0.05, *** P < 0.001, and **** P < 0.0001. Black asterisks represent relative comparison to the “WT control” group, and red asterisks represent comparison to the “mdx control” group. An unpaired two-tailed t test was used for statistical comparisons between mdx-μDys and mdx_μDys + TR groups. t test: * P < 0.05 and ** P < 0.01.

Article Snippet: An anti-MYOM3 polyclonal capture antibody (Proteintech Lab, ref. 17692-1-AP) was coated to the wells of a multiarray plate containing electrodes (MSD, ref. L15XA-3) (overnight, 4°C).

Techniques: Injection, Sequencing, Enzyme-linked Immunosorbent Assay, Muscles, Labeling, Control, Comparison, Two Tailed Test

Journal: Skeletal Muscle

Article Title: Extensive striated muscle damage in a rat model of Duchenne muscular dystrophy with Dmd exons 10–17 duplication

doi: 10.1186/s13395-025-00386-2

Figure Lengend Snippet: Early death associated with severe muscle atrophy in dystrophin-deficient R-DMDdup10-17 (DMD) rats. (A) Scheme of CRISPR/Cas9-mediated duplication on the rat chromosome X of a ~ 125 kb genomic region encompassing exons 10 to 17 of the Dmd gene. The wild-type allele (X + ) is at the top, with the reading frame indicated by the blue colour of the exons. The allele containing the duplication (X DMD ) is at the bottom. Splicing between exon 17 and the duplicated exon 10 induces a premature stop codon in the duplicated exon 10 (white cross on red background) and the absence of translation of subsequent exons (grey). The position and orientation of the primers used for genotyping is indicated by arrows, as is the expected amplicon size for each of the two alleles. (B) Electrophoretic gel of amplicons obtained using a multiplex of the three primers indicated in (A) from genomic DNA of male rats from a litter born from a carrier female and a WT male. A band at 130 bp reveals the WT allele, while a band at 277 bp reveals a male hemizygous for the Dmd allele carrying the duplicated 10–17 region. (C) Body mass of WT and DMD littermates at 6 and 10 months of age. (D) Kaplan-Meier curve for the frequency of WT (black curve) and DMD (orange curve) rat survival. (E) Representative immunofluorescence for laminin (red) and dystrophin (orange) at 6 months on TA, diaphragm and heart sections of WT and DMD littermates, showing complete absence of a dystrophin signal in DMD muscles. Scale bars, 50 μm. (F) Plasma CK levels in 7-month-old WT and DMD littermates. One-tailed unpaired t test. (G) Upper panel: Capillary immunoelectrophoresis plasma protein analysis detecting the two fragments (100 and 130 kDa) of the sarcomeric myomesin-3 (MYOM3) protein in 7-month-old DMD rats, but not in their WT littermates; lower panel: total proteins for each lane. (H) Quantification of plasma MYOM3 levels assessed in ( G ), corresponding to the sum of both MYOM3-130 and MYOM3-100 signals normalised to total plasma proteins. One-tailed unpaired t test. (I) Plasma hs-cTNT levels in 10-month-old WT and DMD littermates. One-tailed unpaired t test

Article Snippet: MYOM3 plasma concentration was obtained with the SimpleProtein Jess analyzer (BioTechne) using the regular 12–230 kDa kit, the polyclonal anti-MYOM3 antibody (ProteinTech #17692-1-AP, 1:2000) and the anti-rabbit HRP (ProteinSimple).

Techniques: CRISPR, Amplification, Multiplex Assay, Immunofluorescence, Muscles, Clinical Proteomics, One-tailed Test, Immunoelectrophoresis

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: International Journal of Molecular Sciences

Article Title: Valproic Acid Improves Antisense-Mediated Exon-Skipping Efficacy in mdx Mice

doi: 10.3390/ijms26062583

Figure Lengend Snippet: Figure 2. Functional recovery following the combined therapy ASO+VPA. (A) Detection of dystrophin protein (green staining) by immunostaining on transverse sections of muscle tissues (triceps and heart) from WT and mdx mice treated with saline, ASO, VPA or ASO+VPA. Nuclei are labelled with DAPI (blue staining). Scale bar, 100 µm. (B) Quantification of the dystrophin intensity staining in heart and triceps; n = 4 mice per group. (C) Myomesin-3 levels in serum of treated mice detected by Western blot; n = 5–7 mice per group. (D) Latency to fall in seconds in the inverted grid test; n = 6–9 mice per group. (E) Latency to fall in seconds in the wire test; n = 6–9 mice per group. (F) Maximal specific force in mg/g measured from the two tibialis anterior muscles of each mouse; n = 6–8 mice per group. (G) percentage of force drop following a series of 15 eccentric contractions measured on semi-isolated tibialis anterior muscles from treated mdx mice; n = 6–8 mice per group. Results are expressed as the mean ± SEM; * p < 0.05, *** p < 0.001, **** p < 0.0001 and ns: non statistically significant analyzed by two-way ANOVA.

Article Snippet: Myomesin-3 was detected by probing the membrane with a primary rabbit polyclonal antibody against MYOM3 (Proteintech, Manchester, UK), followed by a secondary goat anti-rabbit antibody (IRDye 800CW goat anti-rabbit IgG, Li-Cor, Bad Homburg, Germany).

Techniques: Functional Assay, Staining, Immunostaining, Saline, Western Blot, Muscles, Isolation

Journal: The Journal of Clinical Investigation

Article Title: A functional mini-GDE transgene corrects impairment in models of glycogen storage disease type III

doi: 10.1172/JCI172018

Figure Lengend Snippet: ( A ) 4-month-old male Agl –/– mice were injected in the tail vein with an rAAV-MT vector encoding ΔNter2-GDE, at the dose of 1 × 10 14 vg/kg. PBS-injected Agl +/+ and Agl –/– mice were used as controls. ( B ) Western blot analysis of GDE and vinculin expression in heart and triceps, 3 months after vector injection. ( C and D ) Glycogen content measured in heart ( C ) and triceps ( D ) 3 months after vector injection. ( E and F ) Histological analysis of heart ( E ) and triceps ( F ) using HPS and PAS staining. Representative images are shown ( n = 7–9). ( G ) Western blot analysis of Myom3 fragments in plasma of mice 3 months after vector injection. Plasma from mdx mouse was used as positive control. ( H ) Wire-hang test expressed as number of falls per minute, performed before and 3 months after vector injection. Statistical analyses were performed by 1-way ANOVA in C and D and 2-way ANOVA in H .* P < 0.05, ** P < 0.01, *** P < 0.001 versus PBS-injected Agl –/– mice; # P < 0.05, ## P < 0.01, ### P < 0.001 versus PBS-injected Agl +/+ mice. n = 7–9 mice per group coming from 2 independent experiments. All data are shown as mean ±SEM. Scale bars, 50 μm. HPS, hematoxylin phloxine saffron; Myom3, myomesin 3; PAS, periodic acid schiff.

Article Snippet: After transfer, the membrane was blocked with Intercept Blocking buffer (LI-COR Biosciences) and incubated with either an anti-GDE rabbit polyclonal antibody (16582-1-AP, Proteintech for muscles or AS09454, Agrisera for liver; 1:1,000) and an anti-vinculin mouse monoclonal antibody (V9131, Sigma-Aldrich) for tissue lysates and with an anti-Myom3 rabbit polyclonal antibody (17692-1-AP, Proteintech, 1:1,000) for plasma.

Techniques: Injection, Plasmid Preparation, Western Blot, Expressing, Staining, Clinical Proteomics, Positive Control

Journal: The Journal of Clinical Investigation

Article Title: A functional mini-GDE transgene corrects impairment in models of glycogen storage disease type III

doi: 10.1172/JCI172018

Figure Lengend Snippet: ( A ) 6-week-old male Agl –/– rats were injected in the tail vein with an rAAV-MT vector encoding the ΔNter2-GDE, at the dose of 1 × 10 14 vg/kg. PBS-injected Agl –/– and Agl +/+ rats were used as controls. ( B ) Western blot analysis of GDE and vinculin expression in heart and triceps, 3 months after vector injection. ( C and D ) Glycogen content measured in triceps ( C ) and heart ( D ) 3 months after vector injection. ( E and F ) Histological analysis of heart ( E ) and triceps ( F ) with HPS and PAS staining. Representative images are shown ( n = 5). ( G ) Western blot analysis of Myom3 fragments in the plasma of rats 3 months after vector injection. Plasma from mdx mouse was used as positive control. ( H ) Quantification of Myom3 expression on the Western blot showed in Panel G (arbitrary units). Statistical analyses were performed by 1-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001 versus PBS-injected Agl –/– rats; # P < 0.05, ## P < 0.01, ### P < 0.001 versus PBS-injected Agl +/+ rats; n = 5 rats per group. All data are shown as mean ±SEM. Scale bars, 50 μm. HPS, hematoxylin phloxine saffron; Myom3, myomesin 3; PAS, periodic acid schiff.

Article Snippet: After transfer, the membrane was blocked with Intercept Blocking buffer (LI-COR Biosciences) and incubated with either an anti-GDE rabbit polyclonal antibody (16582-1-AP, Proteintech for muscles or AS09454, Agrisera for liver; 1:1,000) and an anti-vinculin mouse monoclonal antibody (V9131, Sigma-Aldrich) for tissue lysates and with an anti-Myom3 rabbit polyclonal antibody (17692-1-AP, Proteintech, 1:1,000) for plasma.

Techniques: Injection, Plasmid Preparation, Western Blot, Expressing, Staining, Clinical Proteomics, Positive Control