We used Abs against the C-terminal region of rat and mouse LAMP-1 (ab24170; Abcam, Cambridge, MA), LAMP-2 (L0668; Sigma-Aldrich, St. Louis, MO), LAMP-2a (51-2200; Invitrogen/Life Technologies, Carlsbad, CA), and lysosomal integral membrane protein type-2 (LIMP-2) (NB400-129; Novus Biologicals Oakville, ON, Canada); and against full-length/luminal mouse LAMP-1 (1d4b; Iowa Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and mouse LAMP-2 (GL2A7; Iowa Developmental Studies Hybridoma Bank). The following Abs were used for immunofluorescence and immunohistochemical analyses: collagen-1 (ab292; Abcam), Ki67 (IHC-00375; Bethyl Laboratories, Inc. Montgomery, TX), α-smooth muscle actin (α-SMA) (clone 1A4; Sigma-Aldrich), insulin (4590; Cell Signaling Technology, Beverly, MA), CD68 (ABIN181836; Antibodies Online, Atlanta, GA), and CD206 (OASA05048; Aviva Systems Biology, San Diego, CA).
α smooth muscle actin α sma clone 1a4
Manufactured by Merck Group
Sourced in Japan, Germany
α-smooth muscle actin (α-SMA) (clone 1A4) is a laboratory tool used to identify and quantify smooth muscle cells. It is a protein marker that is expressed in smooth muscle cells and is commonly used in research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab292, by Abcam (1 mentions)
Ab24170, by Abcam (1 mentions)
L0668, by Merck Group (1 mentions)
Nb400 129, by Novus Biologicals (1 mentions)
N histofine simple stain rat max po, by Nichirei Biosciences (1 mentions)
Vectastain abc ap, by Vector Laboratories (1 mentions)
Irdye 680 and 800, by LI COR (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Anti tubulin clone dm1a, by Merck Group (1 mentions)
Bond max, by Leica (1 mentions)
Alexa fluor 594 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)
Clone mec 13, by BD (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Cy3 conjugated mouse monoclonal anti α smooth muscle actin antibody, by Merck Group (1 mentions)
Rabbit anti mouse ng2 antibody, by Merck Group (1 mentions)
4 protocols using α smooth muscle actin α sma clone 1a4
1
Immunohistochemical Profiling of Lysosomal Markers
2
Histological Analysis of Vascular Graft Implantation
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The implanted SF graft was hollowed out together with the skin with surgical scissors and divided in half. In addition, the central part of the graft was cut 4 mm transversely, and the sutured part of the remaining native blood vessel and artificial vascular graft was cut longitudinally. These samples were fixed with ethanol for histological analyses. The fixed samples were embedded in paraffin and processed for hematoxylin and eosin (H&E) and Masson’s trichrome (MTC) staining. Sections for immunohistochemical staining were incubated with primary antibodies, including α-smooth muscle actin (α-SMA; clone 1A4; Sigma-Aldrich, Inc., Tokyo, Japan) and CD31 anti-rat antibody (BD Biosciences, Inc., Tokyo, Japan). The A-SMA sample was incubated with biotinylated anti-mouse immunoglobulin secondary antibody (Vector Laboratories, Inc., Burlingame, CA, USA), and subsequent color development was obtained using Vectastain ABC-AP (Vector Laboratories, Inc., Burlingame, CA, USA). The CD31 sample was incubated with N-Histofine Simple Stain Rat MAX PO (Nichirei Biosciences, Inc., Tokyo, Japan), and subsequent color development was obtained using Vectastain ABC-AP (Vector Laboratories, Inc., Burlingame, CA, USA).
3
Western Blot Analysis of αSMA in NAFs and CAFs
4
Immunohistochemical Analysis of Murine Tumour Tissue
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Murine tumour tissue samples were snap-frozen and prepared for microscopical analyses as described (Coutelle et al, 2015 (link)). As primary antibody for staining of endothelial cells, pericytes and type IV collagen, rat monoclonal anti-mouse CD31 antibody (PEACAM-1; 1 : 50; clone MEC 13.3, BD Pharmingen, Heidelberg, Germany), Cy3-conjugated mouse monoclonal anti-α-smooth muscle actin antibody (1 : 100; α-smooth-muscle-actin (α-SMA) clone 1 A4, Sigma-Aldrich, Munich, Germany), rabbit anti-mouse NG2 antibody (1 : 200; Millipore, Darmstadt, Germany) and rabbit polyclonal anti-type IV collagen antibody (1 : 100; Clone ab6586, Abcam, Cambridge, UK) were used, respectively. The CD31 antibody was detected by Alexa Fluor 488-conjugated polyclonal goat anti-rat antibody and the NG2 and collagen IV antibody were detected by Alexa Fluor 594-conjugated goat anti-rabbit antibody (1 : 500; Molecular Probes, Eugene, OR, USA). Immunohistochemical staining of human paraffin-embedded tumour sections was performed using the BOND MAX from Leica (Wetzlar, Germany) according to the protocol of the manufacturers. As a primary antibody Collagen IV clone CIV 22 mouse monoclonal, Dako (M0785; Glostrup, Denmark) at a dilution of 1 : 50 was used.
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