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Rabbit anti α smooth muscle actin

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Rabbit anti-α-smooth muscle actin is an antibody that recognizes the α-smooth muscle actin protein. This protein is a component of the contractile apparatus in smooth muscle cells.

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Lab products found in correlation

Mouse anti vimentin, by Merck Group (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions) Cy3 donkey anti rabbit igg, by Jackson ImmunoResearch (2 mentions) Mouse anti zo 1, by Thermo Fisher Scientific (2 mentions) Alexa 488 donkey anti mouse igg, by Thermo Fisher Scientific (2 mentions) Ctr6000, by Leica (2 mentions) Rabbit anti cd31, by Abcam (2 mentions) Lsm 780, by Zeiss (2 mentions) Citrate buffer, by Agilent Technologies (1 mentions) Alexa fluor 647, by Abcam (1 mentions) Alexa fluor 488, by Abcam (1 mentions) Rabbit anti ve cadherin, by Abcam (1 mentions) Fluoview 1000, by Olympus (1 mentions) Rabbit anti ki67, by Thermo Fisher Scientific (1 mentions) Rat anti cd31, by BD (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Mouse anti gfp, by Thermo Fisher Scientific (1 mentions) Rabbit anti cd206, by Abcam (1 mentions) Dapi solution, by Vector Laboratories (1 mentions)

Close spelling variants of this product

Rabbit anti-human alpha-smooth muscle actin (α-SMA) Rabbit anti-α-smooth muscle actin (SMA, Rabbit anti-α-smooth muscle actin (α-SMA Rabbit anti-alpha smooth muscle actin (α-SMA) Rabbit anti-human α-smooth muscle actin (α-SMA) Rabbit anti-alpha smooth muscle actin (α-SMA) antibody Rabbit anti–α‐smooth muscle actin antibody Anti-α-smooth muscle actin α-smooth muscle actin Anti-α-actin

10 protocols using rabbit anti α smooth muscle actin

1

Immunohistochemical Analysis of Penile Tissues

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After fixation, penile tissues were dehydrated and embedded in paraffin. Sections (5μm) from the middle of the body were used for immunohistochemical staining. In brief, the sections were dewaxed and rehydrated in graded ethanol. After heat-induced epitope retrieval with citrate buffer (Dako, Carpinteria, CA), slides were treated with 0.3% hydrogen peroxide in methanol to quench endogenous peroxidase. The sections were then incubated with blocking buffer for 30 min at room temperature. Primary antibody (rabbit anti-α-smooth muscle actin, Abcam, Cambridge, MA; or purified mouse anti-nNOS, BD Biosciences, San Jose, CA) in 1% BSA was applied overnight at 4ºC. After rinsing 3×5 min., the sections were incubated with secondary antibody (biotinylated anti-rabbit or anti-mouse IgG H+L, Vector, Burlingame, CA) for 2 hours at room temperature. The Avidin-Biotin Complex (ABC) staining method was applied. The sections were then counterstained with hematoxylin. No primary antibody control was used to support the specificity of the immunoreactive staining.
Tao M., Tasdemir C., Tasdemir S., Shahabi A, & Liu G. (2017). Penile alterations at early stage of type 1 diabetes in rats. International Brazilian Journal of Urology : official journal of the Brazilian Society of Urology, 43(4), 753-761.
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2

Immunostaining of Cellular Markers

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Immunostaining was performed as previously described29 using the following primary antibodies: mouse anti-CTNT (Thermo NeoMarkers; 1:100), rabbit anti-WT1 (Abcam Epitomics, 1:100), mouse anti-ZO1 (Invitrogen, 1:100), rabbit anti-α-Smooth Muscle Actin (Abcam, 1:100) and mouse anti-Vimentin (Sigma-Aldrich, 1:100). Secondary antibodies used were: donkey anti-rabbit IgGCy3 (Jackson ImmunoResearch; 1:300), donkey anti-mouse IgG-Alexa 488 (Invitrogen; 1:300). DAPI (Life Technologies, SlowFade Gold) was used to counterstain nuclei. The stained cells were visualized using a fluorescence microscope (Leica CTR6000) and images captured using the Leica Application Suite software.
Witty A.D., Mihic A., Tam R.Y., Fisher S.A., Mikryukov A., Shoichet M.S., Li R.K., Kattman S.J, & Keller G. (2014). The generation of the epicardial lineage from human pluripotent stem cells. Nature biotechnology, 32(10), 1026-1035.
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3

Immunostaining of Cellular Markers

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Immunostaining was performed as previously described29 using the following primary antibodies: mouse anti-CTNT (Thermo NeoMarkers; 1:100), rabbit anti-WT1 (Abcam Epitomics, 1:100), mouse anti-ZO1 (Invitrogen, 1:100), rabbit anti-α-Smooth Muscle Actin (Abcam, 1:100) and mouse anti-Vimentin (Sigma-Aldrich, 1:100). Secondary antibodies used were: donkey anti-rabbit IgGCy3 (Jackson ImmunoResearch; 1:300), donkey anti-mouse IgG-Alexa 488 (Invitrogen; 1:300). DAPI (Life Technologies, SlowFade Gold) was used to counterstain nuclei. The stained cells were visualized using a fluorescence microscope (Leica CTR6000) and images captured using the Leica Application Suite software.
Witty A.D., Mihic A., Tam R.Y., Fisher S.A., Mikryukov A., Shoichet M.S., Li R.K., Kattman S.J, & Keller G. (2014). The generation of the epicardial lineage from human pluripotent stem cells. Nature biotechnology, 32(10), 1026-1035.
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4

Immunohistochemical Characterization of InVADE Tissues

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InVADE tissues were fixed after 8 days in culture (4% PFA, 4°C, overnight), then blocked with 10% horse serum and 0.1% Triton-X 100 (1 h, RT). Immunostaining was performed with primary antibodies: rabbit anti-α-smooth muscle actin (Abcam, 1:200, 1 h, RT) and secondary antibody donkey anti-rabbit Alexa Fluor 488 (Abcam; 1:400). Conjugated antibodies of Alexa Fluor 594 anti-cytokeratin 19 (BioLegend; 1:200) were used to stain for CK19. Conjugated vimentin-Cy3 (Sigma; 1:200) was used to stain fibroblasts phenotype prior to use in tissue. Endothelial lining was immunostained with primary antibody rabbit anti-VE-cadherin (Abcam, 1:200, 2h, RT) and secondary antibody donkey anti-rabbit Alexa Fluor 647 (Abcam, 1:200) or primary antibody mouse anti-CD31 (Abcam, 1:200, 2h, RT) and secondary antibody goat anti-mouse Alexa Fluor 647 (Abcam, 1:200). DAPI (Invitrogen, 1:1000) was used as a counterstain for each group. Confocal microscopy images were obtained using an Olympus FluoView 1000 laser scanning confocal microscope (Olympus Corporation) at 20X objectives with imaging parameters kept constant for each region.
Lai Benjamin F.L., Lu Rick X., Hu Y., Davenport H.L., Dou W., Wang E.Y., Radulovich N., Tsao M.S., Sun Y, & Radisic M. (2020). Recapitulating pancreatic tumor microenvironment through synergistic use of patient organoids and organ-on-a-chip vasculature. Advanced functional materials, 30(48), 2000545.
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5

Immunofluorescence Staining of Bladder Tissue

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Sections were fixed with 4% PFA for 5 min followed by 1XPBS washes. Sections were permeabilized with 1XPBS containing 0.1% Triton-X-100 (PBST) for 20 min at room temperature. After blocking with 10% serum containing 4% BSA for 1 h at room temperature in a humidity chamber, the sections were incubated overnight at 4°C with primary antibody: Rabbit anti - Ki-67 (1:250, Thermo Fisher Scientific, Waltham, MA), Goat polyclonal anti - uroplakin II (UPKII, 1:50, Santa Cruz Biotechnology Inc. Dallas, TX); Rat anti - CD31 (1:50, cat. 550274, BD Biosciences, Franklin Lakes, NJ) or Rabbit anti - α smooth muscle actin (1:50, Abcam, Cambridge MA). Only blocking buffer was added to control sections. Sections were washed the next day with PBST four times and incubated with species matched Alexa Fluor 488 or 555 conjugated secondary antibody for 1 h at room temperature and counterstained with DAPI (Sigma, St. Louis, MO). Sections were again washed with PBST three times and mounted with ProLong Gold antifade reagent (Life Technologies, Carlsbad, CA).
Mann E.A., Alam Z., Hufgard J.R., Mogle M., Williams M.T., Vorhees C.V, & Reddy P. (2015). Chronic Social Defeat, But Not Restraint Stress, Alters Bladder Function in Mice. Physiology & behavior, 150, 83-92.
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6

Comprehensive Immunohistochemical Analysis of Heart Tissue

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Immunohistochemistry was performed as described previously31 (link)–33 (link). Briefly, heart tissue sections were permeabilized with PBS containing 0.5% Triton X-100 for 15 min at room temperature. Tissue sections were then blocked with 1% BSA in PBS for 60 min at room temperature and incubated with primary antibodies at 4 ℃ overnight. On the next day, samples were washed three times with PBS and incubated with secondary antibodies for 1h at room temperature. DAPI solution (VectaShield) was added for nuclear staining. The primary antibodies used in this study included mouse anti-Vimentin (Millipore; 1:100), rabbit anti-α smooth muscle actin (Abcam; 1:100), anti-cardiac Troponin T (Thermo; 1:100), rabbit anti-CD31 (Abcam; 1:100), mouse anti-CD68 (Abcam; 1:200), rabbit anti-CD206 (Abcam; 1:200), mouse anti-α sarcomeric actinin (Sigma; 1:100), and mouse anti-GFP (Thermo; 1:100). Secondary antibodies used in this study included either anti-mouse/-rabbit IgG Alexa Fluor 488 (Invitrogen; 1:500) or anti-mouse/-rabbit IgG Alexa Fluor 568 (Invitrogen; 1:500). Imaging of heart sections was performed using a Laser Scanning Microscope LSM 880 NLO with Airyscan (Zeiss).
Lee S., Lee D.H., Park B.W., Kim R., Hoang A.D., Woo S.K., Xiong W., Lee Y.J., Ban K, & Park H.J. (2019). In vivo transduction of ETV2 improves cardiac function and induces vascular regeneration following myocardial infarction. Experimental & Molecular Medicine, 51(2), 1-14.
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7

Comprehensive Antibody Characterization for Western Blotting and Immunostaining

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Primary antibodies used for Western blotting and immunostaining were chicken anti-HMGCS2 (Genway, San Diego, CA), rabbit anti-HMGCS2 (AVIVA, San Diego, CA; epitope different from the Genway antibody), chicken anti-MCT1 (Chemicon, Temecula, CA), rabbit anti-MCT1, goat anti-PPARα (Santa Cruz Biotech­nology, Dallas, TX), rabbit anti–α-smooth muscle actin, rabbit anti-prohibitin (Abcam, Cambridge, UK), rabbit anti–cytochrome c, rabbit anti-Glut4, rabbit anti–COX IV, rabbit anti-ACC, rabbit anti–phospho-ACC, rabbit anti-AMPKα, rabbit anti–P-AMPKα, rabbit anti-AMPKβ1, rabbit anti–P-AMPKβ1 (Cell Signaling Technology, Danvers, MA), rat anti-Hsc70 (Enzo Life Sciences, Helsinki, Finland), rat anti-K8 (Troma I Developmental Studies Hybridoma Bank, Iowa City, IA), and mouse anti-tubulin (Sigma-Aldrich, Munich, Germany). Secondary antibodies used for Western blotting were anti-mouse immunoglobulin G (IgG)–horseradish peroxidase (HRP), anti-rat IgG-HRP (GE Healthcare, Buckinghamshire, UK), anti-rabbit IgG-HRP (Cell Signaling Technology), anti-chicken IgG-HRP (Genway), and anti-goat IgG-HRP (Cell Signaling Technology). Secondary antibodies used for immunostaining were anti-rabbit Alexa Fluor 488 and anti-rat Alexa Fluor 546 (Life Technologies, Carlsbad, CA). Nuclei were stained with Toto-3 (Life Technologies) or DRAQ5 (Cell Signaling Technology).
Helenius T.O., Misiorek J.O., Nyström J.H., Fortelius L.E., Habtezion A., Liao J., Asghar M.N., Zhang H., Azhar S., Omary M.B, & Toivola D.M. (2015). Keratin 8 absence down-regulates colonocyte HMGCS2 and modulates colonic ketogenesis and energy metabolism. Molecular Biology of the Cell, 26(12), 2298-2310.
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8

Protein Expression Analysis of Left Ventricle

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Tissue homogenates were prepared by placing left ventricle tissue in ice-cold RIPA lysis buffer (pH 7.0) from Santa Cruz Biotechnology (Santa Cruz, CA; SC-24948) containing protease inhibitors and phosphatase inhibitors. About 40 µg homogenates were analyzed using Western blot. The primary antibodies used in western blot analyses were: Rabbit anti-CD31, 1:500 dilution (Abcam Inc., Cat. No.: ab28364); Rabbit anti-α-smooth muscle actin, 1:500 dilution (Abcam Inc., Cat. No.: ab5694); Rabbit anti-GAPDH antibody, 1:1000 dilution (Santa Cruz Inc., Cat. No.: sc-25778). Secondary antibodies included goat anti-rabbit IgG-HRP (Santa Cruz Inc., Cat. No.: sc-2030); goat anti-mouse IgG-HRP (Santa Cruz Inc., Cat. No.: sc-2031). The Western blot images were cropped for presentation in each figure, and the uncropped Western blot images were provided in the Supplementary Info File.
Shi H., Fan X., Horton A., Haller S.T., Kennedy D.J., Schiefer I.T., Dworkin L., Cooper C.J, & Tian J. (2019). The Effect of Electronic-Cigarette Vaping on Cardiac Function and Angiogenesis in Mice. Scientific Reports, 9, 4085.
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9

Immunofluorescence Analysis of Airway Tissue

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After the process of deparaffinization and antigen retrieval as described above, tissue sections were permeabilized with 0.1% Tween 20 in PBS. Tissue sections were blocked with blocking reagent and incubated overnight at 4 °C with rabbit anti-α-smooth muscle actin (1:500; Abcam, Cambridge, MA), mouse anti-PGP 9.5 (1:100; Abcam), rabbit anti-von Willebrand factor (1:200; Dako) and mouse anti-D2-40 (1:200; Dako) followed by donkey anti-rabbit Alexa fluor 488 antibody or goat anti-mouse Alexafluor 555 antibody (Invitrogen, Burlington, ONT) for 1 h. Diluent without primary antibodies was used as control. After washing with PBS, cell nucleus was stained with Hoechst 33342 (AnaSpec, Fremont, CA) for 10 min and mounted with PermaFluor (Thermo scientific, Fremont, CA). Images were captured with a confocal microscope (LSM780; Carl Zeiss Microscopy, Jena, Germany) and analyzed using ImageJ (National Institutes of Health, Bethesda, MD) software. ASM and nerve area were expressed as percentage of α-ASMor PGP 9.5-positive area over whole airway area in the section, calculated using ImageJ software.
Ichikawa T., Panariti A., Audusseau S., Mogas A.K., Olivenstein R., Chakir J., Laviolette M., Allakhverdi Z., Al Heialy S., Martin J.G, & Hamid Q. (2019). Effect of bronchial thermoplasty on structural changes and inflammatory mediators in the airways of subjects with severe asthma. Respiratory medicine, 150.
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10

Analyzing Murine Kidney Protein Expression

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Murine kidney tissues were lysed with ice-cold lysis buffer (Thermo Fisher Scientific) supplemented with complete protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific). After centrifugation at 10000 rpm for 5 min, proteins were boiled in Roti-Load1 Buffer (Carl Roth) at 100ºC for 5 min. Proteins were separated on SDS-polyacrylamide gels and transferred to PVDF membranes. The membranes were incubated overnight at 4°C with the following primary antibodies: rabbit anti-α-smooth muscle actin, rabbit anti-collagen I (diluted 1:1000, Abcam), mouse anti-fibronectin (diluted 1:1000, BD Biosciences), rabbit anti-Tak1, rabbit anti-Smad2, rabbit anti-phosphorylated Smad2 (Ser 465/467 , diluted 1:1000, Cell Signaling),
Feger M., Alesutan I., Castor T., Mia S., Musculus K., Voelkl J, & Lang F. (2015). Inhibitory effect of NH4Cl treatment on renal Tgfß1 signaling following unilateral ureteral obstruction. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 37(3).
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