antibodies against myhc  (Developmental Studies Hybridoma Bank)

Journal: Poultry Science

Article Title: METTL14-mediated RNA m 6 A modifications of TET1 and TET2 in duck myoblast differentiation

doi: 10.1016/j.psj.2026.106435

Figure Lengend Snippet: Comparison of RNA m 6 A modification levels in duck leg muscle. (A) DPMs during the proliferation phase; (B) DPMs on day 2 of differentiation (myotubes); (C) Immunofluorescence for Desmin in DPMs during proliferation; (D) Immunofluorescence for MYHC in DPMs on day 4 of differentiation. Dot blot and quantitative analysis of m 6 A in total RNA from duck embryos at E10 and E20 are shown in panels (E) and (F); panels (G) and (H) display quantitative and dot blot analyses of m 6 A in total RNA from DPMs in the GM and DM phases. Methylene blue (MB) staining served as a control for RNA loading. E10 and E20 indicate leg muscle samples from duck embryos at 10 and 20 days of incubation, respectively. GM and DM represent undifferentiated DPMs and DPMs at 48 hours post-differentiation, respectively. * indicates P < 0.05, ** indicates P < 0.01.

Article Snippet: The membrane was blocked in 5% nonfat milk in PBS for 2 hours, followed by overnight incubation at 4°C with primary antibodies against MYHC (DSHB, Iowa, IA, USA; 0.3 μg/mL) and GAPDH (Proteintech, Wuhan, China; 1:50000).

Techniques: Comparison, Modification, Immunofluorescence, Dot Blot, Staining, Control, Incubation

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: Cross-sections (10 µm) of bovine longissimus dorsi muscle biopsy samples were stained with a primary antibody against laminin (ab7463, Rabbit, 1:100) to label the myofiber boundaries, together with a cocktail of mouse monoclonal antibodies against MyHC type I (BA-D5c, MIgG2b. 1:100), MyHC type IIA (SC-71c, MIgG1, 1:100), and MyHC type IIB (BF-F3c, MIgM, 1:100). Primary antibody staining was visualized using Alexa Fluor-conjugated secondary antibodies (1:500) including Goat Anti-Rabbit Alexa Fluor 350 (to detect laminin), Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect SC-71), Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5), and Goat Anti-Mouse IgM Alexa Fluor 647 (to detect BF-F3). Laminin, BA-D5, SC-71, and BF-F3 staining were pseudo colored white, red, green, and blue respectively. Scale bars 100 µm.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining, Bioprocessing

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: A: Cross-sections (10 µm) of bovine longissimus dorsi muscle biopsy samples were stained with a primary antibody against dystrophin [MANDYS1(3B7)s, MIgG2a, 1:10] to label the label the myofiber boundaries, together with a cocktail of mouse monoclonal primary antibodies against MyHC type I (BA-D5c, MIgG2b, 1:100) and MyHC type IIA (SC-71c, MIgG1, 1:100). Primary antibody staining was visualized using Alexa Fluor-conjugated secondary antibodies (1: 500) including Goat Anti-Mouse IgG2a Alexa Fluor 647 (to detect dystrophin), Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect SC-71), and Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5). Dystrophin, BA-D5, and SC-71 staining was pseudo colored white, red, and green, respectively. B: Serial cross-sections from the same bovine longissimus dorsi muscle biopsy sample shown in panel A were stained with a primary antibody against dystrophin [MANDYS1(3B7)s, MIgG2a, 1:10] to label the label the myofiber boundaries in combination with a cocktail of primary antibodies against MyHC type IIX (6H1s, MIgM, 1:10) and all MyHC isoforms except for type IIX (BF-35c, MIgG1, 1:100). Primary antibody staining was visualized using Alexa Fluor-conjugated secondary antibodies (1:500) including Goat Anti-Mouse IgG2a Alexa Fluor 647 (to detect dystrophin), Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect BF-35), and Goat Anti-Mouse IgM Alexa Fluor 555 (to detect 6H1). Dystrophin, BF-35, and 6H1 staining were pseudo colored white, red, and green, respectively. Scale bars are 200 µm.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: Cross-sections (10 µm) of bovine longissimus dorsi muscle biopsy samples were stained with a primary antibody against laminin (ab7463, Rabbit, 1:100) to label the myofiber boundaries, together with a cocktail of mouse monoclonal primary antibodies against MyHC type I (BA-D5, MIgG2b), MyHC type IIX (6H1, MIgM), and all MyHC isoforms except type IIX (BF-35, MIgG1). Primary antibody binding was visualized with Alexa Fluor-conjugated secondary antibodies (1:500) including Goat Anti-Rabbit Alexa Fluor 350 (to detect laminin), Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect BF-35), Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5), and Goat Anti-Mouse IgM Alexa Fluor 647 (to detect 6H1). Laminin, BA-D5, BF-35, and 6H1 staining were pseudo colored white, red, green, and blue, respectively. Scale bars are 100 µm.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining, Binding Assay

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: Serial muscle cross-sections (10 µm) of bovine longissimus dorsi muscle biopsy samples were stained with a primary antibody against laminin (ab7463, Rabbit, 1:100) to label the myofiber boundaries together with mouse monoclonal primary antibodies against MyHC type I (BA-D5c, MIgG2b, 1:100), MyHC type IIX (6H1s, MIgM, 1:10), and all MyHC isoforms except type IIX (BF-35c, MIgG1, 1:100), applied either individually or in combination as a cocktail. Primary antibody staining was visualized with a cocktail of Alexa Fluor-conjugated secondary antibodies (1:500) including Goat Anti-Rabbit 350 (to detect laminin), Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5), Goat Anti-Mouse IgM Alexa Fluor 647 (to detect 6H1), and Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect BF-35). Laminin, BA-D5, BF-35, and 6H1 were pseudo colored white, red, green, and blue, respectively. Scale bars are 100 µm.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: A: Cross-sections (10 µm) of bovine longissimus dorsi muscle biopsy samples were stained with a primary antibody against laminin (ab7463, Rabbit, 1:100) to label the myofiber boundaries in combination with a mouse monoclonal primary antibody against MyHC type I (BA-D5c, MIgG2b, 1:100). Primary antibody binding was visualized using Alexa Fluor-conjugated secondary antibodies (1:500) including Goat Anti-Rabbit Alexa Fluor 647 (to detect laminin) and Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5). Laminin and BA-D5 staining was pseudo colored white and red, respectively. Stitched panoramic images of entire muscle biopsy cross-sections were obtained using an automated fluorescent microscope (Echo Revolution) and analyzed using the MuscleJ 1.0.2 plugin for FIJI/ImageJ with minor custom modification. Segmented myofiber regions of interest (ROIs) were overlaid on the original images and corresponding cartography maps of type I fibers (red) versus non-type I (e.g., type II) fibers (gray) are shown. Scale bars are 400 µm on stitched mosaic images and 100 µm on representative fields of view. B: Comparison of total myofiber count data between manual image analysis and MuscleJ. C: Comparison of the percentage (%) composition of type I (BA-D5 pos ) fibers between manual image analysis and MuscleJ at different sensitivity thresholds for type I fiber identification. D: Comparison of the percentage (%) composition of type II myofibers (BA-D5 neg ) fibers between manual image analysis and MuscleJ at different sensitivity thresholds for type I fiber identification. B-D: Data is presented as the mean ± SEM of muscle biopsy samples from n=13 animals. ** and **** denote p<0.01 and p<0.0001 between the indicated groups, respectively.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining, Binding Assay, Microscopy, Modification, Comparison

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: Bovine longissimus dorsi muscle biopsy cross-sections were stained with a primary antibody against laminin (ab7463, Rabbit, 1:100) to label the myofiber boundaries together with a cocktail of mouse monoclonal primary antibodies against MyHC type I (BA-D5c, MIgG2b, 1:100), all but IIX (BF-35c, MIgG1, 1:100), and type IIX (6H1s, IgM, 1:10). Primary antibody binding was detected using Alexa Fluor-conjugated secondary antibodies (1:500) including Goat Anti-Rabbit Alexa Fluor 350 (to detect laminin), Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5), Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect BF-35), and Goat Anti-Mouse IgM Alexa Fluor 647 (to detect 6H1). Laminin, BA-D5, SC-71, and 6H1 staining was pseudo colored white, red, green, and blue, respectively. Stitched panoramic images of the entire muscle biopsy cross-section were obtained using an automated fluorescent microscope (Echo Revolution) and analyzed using the MuscleJ plugin for FIJI/ImageJ with custom modifications. Original images and fiber type cartography maps of type I (red), type IIA (green), and type IIX (gray) are shown. Scale bars are 400 µm on stitched mosaic images and 100 µm on representative fields of view.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining, Binding Assay, Microscopy

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: A: Bovine longissimus dorsi muscle cross-section (10 µm) were stained with a primary antibody against laminin (ab7463, Rabbit, 1:100) to label the myofiber boundaries together with a cocktail of mouse monoclonal antibodies against MyHC type I (BA-D5c, MIgG2b, 1:100), MyHC type IIX (6H1s, MIgM, 1:10), and all MyHC types except IIX (BF-35c, MIgG1, 1:100). Primary antibody staining was visualized with Alexa Fluor-conjugated secondary antibodies (1:500) including Goat Anti-Rabbit Alexa Fluor 350 (to detect laminin), Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5), Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect BF-35), and Goat Anti-Mouse IgM Alexa Fluor 647 (to detect 6H1). Laminin, BA-D5, BF-35, and 6H1 were pseudo colored white, red, green, and blue, respectively. Stitched panoramic images of the entire muscle biopsy cross-section were obtained using an automated fluorescent microscope (Echo Revolution) and analyzed using the MuscleBos plugin for FIJI/ImageJ. An original image and corresponding manually or MuscleBos generated fiber type cartography maps are shown. Scale bars on representative fields of view are 100 µm. B–E: Comparison of manual and MuscleBos analysis of the same bovine muscle cross-sections for total myofiber count ( B ), the percentage of type I myofibers ( C ), the percentage of type IIA myofibers ( D ), and the percentage of type IIX myofibers ( E ). F–H: Comparison of manual and MuscleBos analysis of the same bovine muscle cross-sections for mean myofiber cross-sectional area (CSA) measurements for all fibers irrespective of type ( F ), type I fibers ( G ), type IIA fibers ( H ), and type IIX fibers ( I ). Dot plots represent data from each individual animal (biological replicates) for n=13 cows. NS denotes no significant difference between manual analysis and MuscleBos. * and ** denote p<0.05 and p<0.01 between the indicated groups, respectively.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining, Bioprocessing, Microscopy, Generated, Comparison

Journal: bioRxiv

Article Title: Rapid High-Throughput Analysis of Bovine Skeletal Muscle Fiber Morphology via Automated Fluorescent Microscopy and MuscleBos software

doi: 10.64898/2025.12.18.695234

Figure Lengend Snippet: (A): Muscle biopsy samples were obtained from the longissimus dorsi of high muscle (HM) or low muscle (LM) multiparous dairy cows at 21-days before expected calving (PRE) and at 21-days postpartum (POST). Biopsy cross-sections (10 µm) were stained with a primary antibody against laminin (ab7463, Rabbit, 1:100) to label the myofiber boundaries together with a cocktail of mouse monoclonal antibodies against MyHC type I (BA-D5c, MIgG2b, 1:100), MyHC type IIX (6H1s, MIgM, 1:10), and all MyHC types except IIX (BF-35c, MIgG1, 1:10). Primary antibody staining was visualized with Alexa Fluor conjugated secondary antibodies (1:500) including Goat Anti-Rabbit Alexa Fluor 350 (to detect laminin), Goat Anti-Mouse IgG2b Alexa Fluor 555 (to detect BA-D5), Goat Anti-Mouse IgG1 Alexa Fluor 488 (to detect BF-35), and Goat Anti-Mouse IgM Alexa Fluor 647 (to detect 6H1). Laminin, BA-D5, BF-35, and 6H1 were pseudo colored white, red, green, and blue, respectively. Scale bars are 100 µm. B-J: Stitched panoramic images of the entire muscle biopsy cross-section were obtained using an automated fluorescent microscope (Echo Revolution) and analyzed using the MuscleBos plugin for FIJI/ImageJ. B: The overall fiber type profile of pooled bovine longissimus dorsi muscle biopsy samples showing the percentage composition of type I, type IIA, and type IIX fibers as determined by MuscleBos. C: The mean cross-sectional area (CSA) type I, type IIA, and type IIX myofibers in pooled bovine longissimus dorsi muscle biopsy samples as determined by MuscleBos. D: Comparison of mean myofiber CSA irrespective of fiber type between bovine longissimus dorsi muscle biopsy samples obtained from HM or LM cows at PRE and POST parturition. E-G : Comparison of the mean CSA of type I (E) , type IIA (F) , and type IIX (G) myofibers in bovine longissimus dorsi muscle biopsy samples obtained from HM or LM cows at PRE and POST parturition H-J : Comparison of percentage fiber type composition of type I (H) , type IIA (I) , and type IIX (J) myofibers in bovine longissimus dorsi muscle biopsy samples obtained from HM or LM cows PRE and POST parturition. Values are mean ± SEM with dot plots representing data obtained from each individual animal (biological replicates). Symbols denote *p<0.05, **p<0.01, and ***p<0.001, and ****p<0.0001 between the indicated groups.

Article Snippet: Primary antibodies tested included those against dystrophin [Developmental Studies Hybridoma Bank (DSHB), MANDYS1(3B7)s, MIgG2a, 1:10], Laminin 1 + 2 (Abcam, ab7463, Rabbit, 1:100), MyHC type I (DSHB, BA-D5c, MIgG2b, 1:100), MyHC type IIA (DSHB, SC-71c, MIgG1, 1:100), MyHC type IIX (DSHB, 6H1s, MIgM, 1:10), and all but type MyHC IIX (DSHB, BF-35c, MIgG1, 1:100).

Techniques: Staining, Bioprocessing, Microscopy, Comparison