Anti-hypoxia-inducible factor (HIF)-1α and anti-aldehyde dehydrogenase (ALDH) 1 antibodies were purchased from BD Biosciences (San Jose, CA, USA). Anti-CD34, anti-CD44s, anti-glial fibrillary acid protein (GFAP), and anti-human smooth muscle actin (SMA) antibodies were obtained from Dako (Copenhagen, Denmark). Anti-Snail and anti-Slug antibodies were from Cell Signaling Technology (Danvers, MA, USA). Anti-Sox2, anti-S100A4 (rabbit), anti-Twist 1, anti-Nestin, and anti-CD44v6 antibodies were obtained from Abcam (Cambridge, MA, USA). Anti-S100A4 (mouse), anti-carbonic anhydrase (CA) 9, and anti-NMIIA antibodies were from Proteintech (Rosemont, IL, USA). Anti-ZEB 1 and anti-β-actin antibodies were from Sigma-Aldrich Chemicals (St Louis, MA, USA). Blebbistatin and cobalt chloride (CoCl2) were purchased from Toronto Research Chemicals (North York, ON, Canada) and Sigma Chemical, respectively.
Anti human smooth muscle actin
Manufactured by Agilent Technologies
Sourced in Denmark
Anti-human smooth muscle actin (SMA) is a primary antibody that specifically recognizes smooth muscle actin, a key structural protein found in smooth muscle cells. It is used to identify and characterize smooth muscle components in various tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti human cd68, by Agilent Technologies (2 mentions)
Blebbistatin, by LGC (1 mentions)
Anti nestin, by Abcam (1 mentions)
Anti aldehyde dehydrogenase aldh 1, by BD (1 mentions)
Anti glial fibrillary acid protein, by Agilent Technologies (1 mentions)
Anti cd44v6, by Abcam (1 mentions)
Anti zeb1 and anti β actin antibodies, by Merck Group (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Anti tra 1 60, by Merck Group (1 mentions)
Biorevo bz 9000 fluorescence microscope, by Keyence (1 mentions)
Anti ssea4, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Dojindo Laboratories (1 mentions)
Fitc labeling kit nh2, by Dojindo Laboratories (1 mentions)
Anti tra 1 81, by Merck Group (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Anti human nanog, by Cell Signaling Technology (1 mentions)
Anti oct3 4, by Santa Cruz Biotechnology (1 mentions)
Anti α fetoprotein, by R&D Systems (1 mentions)
Alexa fluor 594 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Merck Group (1 mentions)
6 protocols using anti human smooth muscle actin
1
Antibody Characterization for Hypoxia Study
2
Immunocytochemical Characterization of Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunocytochemical analysis was performed as described previously [16 (link)–18 (link)]. The following primary antibodies were used in this study: anti-SSEA4 (1:300 dilution; Millipore), anti-TRA-1-60 (1:300 dilution; Millipore), anti-TRA-1-81 (1:300 dilution; Millipore), anti-Oct-3/4 (1:300 dilution; Santa Cruz Biotechnology), anti-human Nanog (1:800 dilution; Cell Signaling Technology), anti-Class IIIβ-tubulin (TUJ1; 1:500; Covance), anti-human smooth muscle actin (SMA; 1:50; DAKO), and anti-α-fetoprotein (1:100; R&D systems). Cells were incubated with a primary antibody diluted in 1% bovine serum albumin (BSA) and 5% serum containing PBS at 4°C overnight. Secondary staining was performed with an appropriate secondary antibody-conjugated to Alexa Fluor 488 or Alexa Fluor 594 (1:300; Life technologies) for 1 h at room temperature. Lectin staining was performed as described previously [19 (link),20 (link)]. Briefly, rBC2LCN (Wako) was fluorescent-labeled using FITC Labeling Kit-NH2 (Dojindo) according to the manufacturer’s instruction. Next, 10 μg/mL of FITC-conjugated rBC2LCN in 1% BSA containing PBS was used for staining of 4% paraformaldehyde-fixed cells for 1 h at room temperature. Cells were counterstained with DAPI (Dojindo). Images were collected with a BIOREVO BZ-9000 fluorescence microscope (Keyence).
3
Xenograft Mouse Model for Tumor Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
One million cells of each cell line were resuspended in 250 μl of PBS and injected subcutaneously into the flanks of NOD/SCID IL-2Rγ−/− mice in the animal experimentation unit of the University of Granada. 5 wk later, the mice developed tumours that were removed, rinsed with PBS, fixed, and embedded in paraffin at the Biobank of Andalusia Public Health System. Tissue sections were cut and processed for haematoxylin and eosin staining or for immunohistochemistry staining with Anti-Human Smooth Muscle Actin (IR611; Dako), Ms Anti-Human Cytokeratin Clone AE1/AE3 (IR053; Dako), and Rb Anti-Human Glial Fibrillary Acidic Protein (IR 624; Dako). Species-specific secondary antibodies conjugated with peroxidase were used (EnVision Flex/HPR SM802; Dako). Immunohistochemistry staining images were quantified measuring the area that was positive for the signal of the antibody. For each staining, 10 images were quantified using Image J, averaged, and tested statistically using t test.
4
Isolation and Characterization of Vascular SMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Vascular SMCs were isolated from the media of the aortic arch of Smad3−/− and Smad3+/+ mice. The tissue was washed with PBS, cut into 5 mm pieces with the luminal side on 0.1% gelatin-coated cell culture dishes and incubated. After 7–10 days, smooth muscle-like cell outgrowth was observed. SMCs were maintained in DMEM (Lonza, Leusden, the Netherlands), supplemented with 10% fetal calf serum (HyClone, Thermo Scientific, Breda, the Netherlands), 100 U/ml penicillin and 100 μg/ml streptomycin (Sigma-Aldrich, Zwijndrecht, the Netherlands). For characterization, subconfluent SMCs and HUVECs were grown on coverslips and fixed in 2% paraformaldehyde. Cells were permeabilized with PBS/Triton (0.1%) and blocked with PBS + (0.5% BSA/0.15% glycine in PBS). Coverslips were incubated overnight with the primary antibody, anti-Human Smooth Muscle Actin (1:1000 mouse, clone 1A4 Dako). After washing, coverslips were incubated with the secondary antibody, alexa fluor 594 goat anti mouse IgG (1:1000, Life technologies). Coverslips were mounted in Vectashield with Dapi (Vector Laboratories). For the proliferation assay, cells were used at passage 5–11. Smad3+/+ and Smad3−/− VSMCs were seeded in triplicate in 6 cm dishes (5000 cells/well) and allowed to attach. The cells were counted with the coulter counter every day for a week.
5
Immunostaining of Arterial Lesions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For each portion, the eight sections cut at 105 μm intervals from each other were fixed in acetone at room temperature and immunostained with specific antibodies. Primary antibodies, such as anti-human smooth muscle actin (smooth muscle cells (SMC); dilution: 1 : 100; Dako Corporation, Glostrup, Denmark), anti-human CD66b (neutrophils; dilution: 1 : 50; Beckman Coulter, Nyon, Switzerland), and anti-human CRP (dilution: 1/100; Sigma-Aldrich, Saint Louis, MI), were used. For detecting macrophages, we used anti-human CD68 (marker of total macrophages; dilution: 1 : 100; Dako Corporation, CA), anti-human CD86 (marker of M1 macrophages; dilution: 1 : 100; GeneTex Inc., Irvine, CA), anti-human HLA-DR (marker of M1 macrophages; dilution: 1 : 100; Dako Corporation), and anti-human CD163 (marker of M2 macrophages; dilution: 1 : 50; AbD Serotec, Oxford, UK) [26 (link)]. Quantifications were performed using MetaMorph 6 software. Data were presented as percentages of stained area on total lesion area.
6
Quantitative Immunohistochemical Analysis of Carotid Plaques
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen upstream and downstream human carotid specimens were serially cut into eight 7 μm sections per each portion separated by 105 μm from each other [18 (link)]. Sections were fixed in acetone and immunostained with specific antibodies, anti-human smooth muscle actin (dilution: 1 : 100; Dako Corporation, Glostrup, Denmark), anti-human CD68 (dilution: 1 : 100; Dako Corporation), anti-human CD66b (dilution: 1 : 50; Beckman Coulter, Nyon, CH), anti-human matrix metalloproteinase (MMP)-9 (dilution: 1 : 250; Sothern Biotech, Birmingham, AL), anti-human OPG (dilution: 1 : 20; R&D Systems), and anti-human RANKL (dilution: 1 : 20; R&D Systems). Quantifications were performed with MetaMorph software. Results for these parameters were calculated and expressed as percentages of stained area on total lesion area or number of infiltrating cells on mm2 of lesion area.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!