Whole-mount immunostainings were performed as previously described (Guris et al., 2001 (link); Relaix et al., 2003 (link)) using the following antibodies: MF20 (1:300) that recognizes muscle myosin heavy chains, developed by D.A. Fischman and obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology Iowa City, IA 52242, and Desmin (Dako, 1:200). Immunostaining on sections were performed as previously described (Esteves de Lima et al., 2014 (link), 2016 ). Embryos were included in gelatin/sucrose and snap-frozen. Sections of 12μm were fixed in PFA 4%, permeabilized with Triton 0.5% for 20 min and blocked for 1 h in 5% BSA IgG-free. Overnight incubation at 4°C was performed with the following antibodies: MF20 (1:300) developed by A. Kawakami was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biology Iowa City, IA 52242, MYOG (Santa Cruz, 1:50), cleaved CASP3 (Cell Signaling, 1:100), Desmin (Dako, 1:200), Laminin (Abcam, 1:1,000), β -GAL (Promega, 1:100) and M-Cadherin (BD Biosciences, 1:100). Alexa conjugated secondary antibodies (Thermo Fisher Scientific, 1:500) were incubated for 1 h at room temperature and nuclei were stained with DAPI (1:5,000) for 10 min.
Desmin
Manufactured by Agilent Technologies
Sourced in United States, Denmark, Germany
Desmin is a lab equipment product manufactured by Agilent Technologies. It is a type of intermediate filament protein that is commonly used in the identification and study of muscle cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
α sma, by Agilent Technologies (7 mentions)
Benchmark xt, by Roche (6 mentions)
Vimentin, by Agilent Technologies (4 mentions)
S 100 protein, by Agilent Technologies (3 mentions)
Cytokeratin ae1 3, by Merck Group (3 mentions)
S100 protein, by Roche (3 mentions)
Desmin, by Roche (3 mentions)
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Factor xiiia, by Cell Marque (2 mentions)
Hmb45, by Agilent Technologies (2 mentions)
Aperio imagescope, by Leica (2 mentions)
Nanozoomer, by Hamamatsu Photonics (2 mentions)
Ultraview universal dab kit, by Roche (2 mentions)
Calretinin, by Biocare Medical (2 mentions)
Ini 1, by BD (2 mentions)
α sma, by Roche (2 mentions)
Biotinylated goat anti mouse igg, by Merck Group (2 mentions)
Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions)
Podocin, by Abcam (2 mentions)
α sma, by Merck Group (2 mentions)
53 protocols using desmin
1
Whole-Mount and Section Immunostaining Protocols
2
Comprehensive Multicolor Flow Cytometry Panel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-CD31 (390), mAbs from ThermoFisher; PB- & BV711-anti-mouse CD45 (30-F11 dilution 1/500), AF488-anti-CD115 (AFS98 dilution 1/300), PE-Cy7-anti-CD31 (390 dilution 1/300), AF488- & AF647-anti-CD54 (YN1/1.7.4 dilution 1/300), APC-anti-CD140b (18A2 dilution 1/300), APC-Cy7-anti-CD115 (AFS98 dilution 1/300), AF647- & PE-Cy7-anti-CD117 (2B8 dilution 1/300), PB-FcεRI (MAR1 dilution 1/300), AF647-anti-IL-17A (TC11-18H10.1 dilution 1/300), AF594-anti-CD4 (GK1.5 dilution 1/300), AF700-anti-CD3 (17A2 dilution 1/300), BV605-anti-CD41 (MWReg30 dilution 1/300), PE-anti-CD49d (R1-2 dilution 1/300), AF647-anti-CD11c (N418 dilution 1/300), Rat IgG1 mAbs from Biolegend; Anti-IL-17RA (G-9 dilution 1/100) from SantaCruz; Desmin (D33 dilution 1/100) were obtained from Dako; Anti- αSMA (1A4 dilution 1/300) from Sigma-Aldrich; anti-mouse CXCL1 (polyclonal dilution 1/100) from R&D systems. Anti-mMCP-1 (RF6.1 dilution 1/100) from ThermoFisher. Anti-MRP14 mAb (dilution 1/500) was a gift form Dr Nancy Hogg (The Francis Crick Institute, UK).
3
Skeletal Muscle Cryosectioning and Immunostaining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cryosections, skeletal muscle was dissected, embedded in gum tragacanth (1% in PBS), and frozen in 2-methylbutane cooled liquid nitrogen. Frozen sections were cut at 10 μm and stained with H&E using routine procedures. Immunohistochemistry was performed by fixation with 1% PFA/PBS; permeabilization with 0.2% Triton X-100 in PBS; blocking with PBS/1% BSA, 1% heat-inactivated goat serum, and 0.025% Tween20; incubation with primary antibody for at least 2 h; incubation with secondary Alexa-488 IgG1 antibody (Invitrogen) for 1 h; and mounting with VectaShield containing DAPI (Vector Laboratories). Anti-mouse myosin (my32, Sigma), desmin (DAKO), and myogenin (F5D, Developmental Studies Hybridoma Bank) antibodies were used at 1:100. Slides were visualized using a Zeiss LSM 780 confocal microscope.
4
Immunohistochemical Profiling of Rare Tumors
5
Clinicopathological Analysis of Resected GISTs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eighty consecutive resected GISTs received within a period of 8 years were included in the study. Clinical findings and follow-up data were retrieved from the hospital records. All cases were reviewed for tumour size, location, histological type, cellularity, presence of atypia, necrosis, mitosis, secondary changes, lymph node and distant metastasis. Risk stratification was done according to Miettinen’s classification (8 (link)). Extraintestinal GIST (EGIST) was defined by a combined approach of radiological, operative, gross and microscopic examination where the bulk of the tumour was outside the gastrointestinal tract and did not show a clear cut connection with the bowel or stomach wall. Immunohistochemistry was performed for CD117 (DAKO, Denmark), DOG1 (Novocastra, United States), CD34 (DAKO, Denmark), SMA (DAKO, Denmark), S100 (DAKO, Denmark), desmin (DAKO, Denmark) and vimentin (DAKO, Denmark).
6
Multimodal Protein Profiling in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the stimulation period, cells were fixed in 4% PFA, stained for specific antigens using primary antibodies against desmin (DAKO), DDR2 (Santa Cruz), NCX (SWANT), P4HB, Vimentin, P‐CaMKII, ADAMTS13 (Abcam), incubated with fluorophore‐conjugated secondary antibodies and visualized on Zeiss LSM 510 Meta and Nikon A1 confocal microscopes. The images were analyzed using ImageJ software (ImageJ 1.46r, NIH, USA).
7
Immunohistochemical Analysis of Molar Villi
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Five-μm-thick formaldehyde-fixed paraffin-embedded tissue sections were transferred onto adhesive slides and dried at 62°C for 30 minutes. Following standard heat epitope retrieval for 30 minutes in EDTA, pH 8.0 in the Benchmark automatic immunostaining device (Ventana Medical System, Tucson, AZ), we incubated the samples with primary antibodies against α-SMA (1:200, DAKO, Glostrup, Denmark), PDGFR-β (1:100, Epitomics, Burlingame, CA, USA), and desmin (1:200, DAKO). The sections were subsequently incubated with biotinylated anti-mouse immunoglobulins, peroxidase-labeled streptavidin (LSAB kit, DAKO), and 3,3'-diaminobenzidine. Harris’s hematoxylin served as the counterstain. Primary antibodies were omitted during staining of negative control samples. Blood vessels in the admixed endometrial tissue served as internal positive controls.
Marker immunoreactivity was compared for blood vessels and stromal cells in both the normal and molar chorionic villi. As moles lack a discernable chorionic plate, no comparative immunoreactivity analyses of this region with the normal chorionic plate (from which primary stem villi project vertically downward) was possible.
The staining intensity of the villous stromal cells was scored from 0 to 3 independently by two pathologists (KRK and COS) where: no reactivity (0), weak (+1), moderate (+2), and strong (+3).
Marker immunoreactivity was compared for blood vessels and stromal cells in both the normal and molar chorionic villi. As moles lack a discernable chorionic plate, no comparative immunoreactivity analyses of this region with the normal chorionic plate (from which primary stem villi project vertically downward) was possible.
The staining intensity of the villous stromal cells was scored from 0 to 3 independently by two pathologists (KRK and COS) where: no reactivity (0), weak (+1), moderate (+2), and strong (+3).
8
Immunofluorescent Analysis of Cytoskeletal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After shortly being washed with cold phosphate buffer saline (PBS), cells were fixed with 4% paraformaldehyde at room temperature for 20 min and subsequently stained with primary antibodies at the following dilutions: α-SMA, (1 : 100, DAKO, Hamburg, Germany), vimentin (1 : 200, DAKO), and desmin (1 : 50, DAKO), followed by fluorescent secondary antibodies (DAKO). The nuclei were then counterstained with bisbenzimide. Photos of eight different areas in each well were taken using a microscope with 100x magnification. A minimum of 500 cells in each experimental group was analyzed (n = 3).
9
Characterization of Infant Myoblast Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All data were performed using a human myoblasts isolated from quadriceps muscle biopsies of 5-day-old infant with no neuromuscular disorders, as previously reported71 (link). In order to prove that this cell is a reliable model, we used two other sources of human myoblast, which were quadriceps muscles from 25 and 60-year-old healthy donors. Myoblasts were cultivated in growth medium (DMEM [Gibco, Carlsbad, CA, USA], 50 µg/ml gentamicin [Gibco], 20% [v/v] fetal bovine serum [FBS, Gibco]) and the myogenic purity of the populations was monitored by immunocytochemistry using desmin (Dako, Golstrup, Denmark) as marker. Differentiation was induced at confluence by replacing the growth medium with serum-free DMEM supplemented with 50 µg/mL gentamycin and 10 µg/mL of human insulin (Sigma Aldrich, St Louis, MO, USA). Myotube formation was assessed by fusion index. The fusion index was calculated as the ratio of the number of nuclei inside myotubes to the number of total nuclei ×100 at day 4 of myogenic differentiation, using three independent fields.
10
Immunohistochemical Analysis of Sarcoma Markers
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!