Human/Mouse/Rat alpha -Smooth Muscle Actin Antibody

Detection of alpha-Smooth Muscle Actin in Human Breast Tumor via seqIF™ Staining on COMET™​ ACTA2 was detected in immersion fixed paraffin-embedded sections of human breast tumor using Mouse Anti-Human/Mouse/Rat alpha-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420) at 0.5ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Mouse IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100).. Specific staining was localized to the cytoplasm of smooth muscle cells. Protocol available in available in COMET™ Panel Builder.​

Detection of Human alpha ‑Smooth Muscle Actin by Western Blot. Western blot shows lysates of human heart tissue and iBJ6 human induced pluripotent stem cell line undifferentiated or cardiac differentiated. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human/ Mouse/Rat a-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for a-Smooth Muscle Actin at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Mouse and Rat alpha ‑Smooth Muscle Actin by Western Blot. Western blot shows lysates of mouse colon tissue, C2C12 mouse myoblast cell line, NIH-3T3 mouse embryonic fibroblast cell line, and rat colon tissue. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat a-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for a-Smooth Muscle Actin at approximately 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

alpha ‑Smooth Muscle Actin in BG01V Human Embryonic Stem Cells. a-Smooth Muscle Actin was detected in immersion fixed BG01V human embryonic stem cells differentiated to cardiomyocytes using Mouse Anti-Human/Mouse/ Rat a-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cytoskeleton. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

alpha -Smooth Muscle Actin in Human Breast Cancer Tissue. a-Smooth Muscle Actin was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human/ Mouse/Rat a-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420) at 8 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human alpha ‑Smooth Muscle Actin by Simple Western^TM. Simple Western lane view shows lysates of iBJ6 human induced pluripotent stem cell line untreated (-) or differentiated to cardiomyocytes (+), loaded at 0.2 mg/mL. A specific band was detected for alpha ‑Smooth Muscle Actin at approximately 49 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat alpha ‑Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

alpha-Smooth Muscle Actin in Human Breast Cancer Tissue Using Dual RNAscope^®ISH and IHC. alpha-Smooth Muscle Actin mRNA was detected (top image) in formalin-fixed paraffin-embedded tissue sections of human breast cancer tissue probed with ACD RNAScope® Probe (Catalog # 444771-C2) and stained using ACD RNAscope® 2.5 HD Duplex Detection Reagents (red, Catalog # 322500). Adjacent tissue section (bottom image) was processed for immunohistochemistry using R&D Systems Mouse Anti-Human/Mouse/Rat alpha-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420) at 0.5 μg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte HRP Polymer Antibody (VC001) and DAB chromogen (brown). Tissues were counterstained with hematoxylin (blue).

Detection of alpha ‑Smooth Muscle Actin in Human Blood Monocytes by Flow Cytometry. Human peripheral blood monocytes were stained with Mouse Anti-Human/Mouse/Rat a-Smooth Muscle Actin Monoclonal Antibody (Catalog # MAB1420, filled histogram) or isotype control antibody (MAB003, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Detection of Human alpha-Smooth Muscle Actin by Immunocytochemistry/Immunofluorescence Overview of Multi-dimensional Microscopic Molecular Profiling (MMMP).The overall MMMP approach is depicted using an example tissue section from normal human duodenum (sample #1.9.7). (a) Slides were subjected to repeated cycles of staining and imaging with fluorescent primary antibodies and DAPI. At the end of each cycle, fluorescent signal was removed by a chemical bleaching process, and slides were again imaged, before proceeding to the next round of this iterative procedure. After the final antibody stain (#15 Sma), slides were analyzed with a series of histochemical stains. (b) A set of tiling images spanning each tissue section was initially generated by the microscope system. The tiling images were then computationally ‘stitched’ together to produce a single image per staining cycle for each sample. (c) Image registration was performed to align images from the same tissue section across cycles. Mean intensities of the DAPI signal from all immuno-fluorescence images are shown from before (Unregistered) and after (Registered) the image registration procedure was completed. (d) Following registration, signal intensities from the relevant channels for each image (columns) in the MMMP series were extracted for each pixel (rows) within the tissue section and compiled into a large data matrix of in situ molecular profiles. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0128975), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human alpha-Smooth Muscle Actin by Immunocytochemistry/Immunofluorescence Functional glycosylation of alpha ‐dystroglycan and characterization of dystroglycanopathy patient‐specific iPSCsCurrent model of the core M3 functional glycan structure on alpha ‐dystroglycan and enzymes involved in its synthesis. ECM ligands, such as laminins, bind to the Xyl‐GlucA disaccharide repeats (IIH6 epitope). Man, mannose; GlcNAc, N‐acetylglucosamine; GalNAc, N‐acetylgalactosamine; Rbo5P, ribitol‐5‐phosphate; Xyl, xylose; GlcA, glucuronic acid.Representative images of immunostaining demonstrate that FKRP‐iPSCs express specific pluripotency‐associated markers, including NANOG, OCT4, Tra‐1‐60, and SSEA4.FKRP‐iPSCs have a normal karyotype.In vitro differentiation of FKRP‐iPSCs to cells representing ectoderm ( beta ‐III Tubulin, Tuj1), mesoderm (SMA, smooth muscle actin), and endoderm (AFP, alpha ‐fetoprotein).Data information: Scale bars, 50 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31566294), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human/Mouse/Rat alpha-Smooth Muscle Actin Antibody by Immunohistochemistry Overview of Multi-dimensional Microscopic Molecular Profiling (MMMP).The overall MMMP approach is depicted using an example tissue section from normal human duodenum (sample #1.9.7). (a) Slides were subjected to repeated cycles of staining and imaging with fluorescent primary antibodies and DAPI. At the end of each cycle, fluorescent signal was removed by a chemical bleaching process, and slides were again imaged, before proceeding to the next round of this iterative procedure. After the final antibody stain (#15 Sma), slides were analyzed with a series of histochemical stains. (b) A set of tiling images spanning each tissue section was initially generated by the microscope system. The tiling images were then computationally ‘stitched’ together to produce a single image per staining cycle for each sample. (c) Image registration was performed to align images from the same tissue section across cycles. Mean intensities of the DAPI signal from all immuno-fluorescence images are shown from before (Unregistered) and after (Registered) the image registration procedure was completed. (d) Following registration, signal intensities from the relevant channels for each image (columns) in the MMMP series were extracted for each pixel (rows) within the tissue section and compiled into a large data matrix of in situ molecular profiles. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26176839), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Human/Mouse/Rat alpha-Smooth Muscle Actin Antibody by Immunocytochemistry/ Immunofluorescence Functional glycosylation of alpha ‐dystroglycan and characterization of dystroglycanopathy patient‐specific iPSCsCurrent model of the core M3 functional glycan structure on alpha ‐dystroglycan and enzymes involved in its synthesis. ECM ligands, such as laminins, bind to the Xyl‐GlucA disaccharide repeats (IIH6 epitope). Man, mannose; GlcNAc, N‐acetylglucosamine; GalNAc, N‐acetylgalactosamine; Rbo5P, ribitol‐5‐phosphate; Xyl, xylose; GlcA, glucuronic acid.Representative images of immunostaining demonstrate that FKRP‐iPSCs express specific pluripotency‐associated markers, including NANOG, OCT4, Tra‐1‐60, and SSEA4.FKRP‐iPSCs have a normal karyotype.In vitro differentiation of FKRP‐iPSCs to cells representing ectoderm ( beta ‐III Tubulin, Tuj1), mesoderm (SMA, smooth muscle actin), and endoderm (AFP, alpha ‐fetoprotein).Data information: Scale bars, 50 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31566294), licensed under a CC-BY license. Not internally tested by R&D Systems.