2-Hydroxyethyl methacrylate (2-HEMA) was supplied by Merck (Darmstadt, Germany); ethylene glycol dimethacrylate (EGDMA), Dulbecco’s Modified Eagle’s Medium (DMEM) and dichlorodimethylsilane were from Sigma-Aldrich (Steinheim, Germany); N-(3-aminopropyl) methacrylamide hydrochloride (APMA) was from Polysciences Inc. (Warrington, PA, USA); D(+)-sucrose (99.7%) and 2,2′-azo-bis(isobutyronitrile) (AIBN) from Acros (Geel, Belgium). The Cy3 Ab Labeling Kit was supplied from Amersham/GE Healthcare (Munich, Germany). The AAV Titration ELISA was from Progen (Heidelberg, Germany) and the IGF-1 ELISA kit (Insuline like Growth Factor 1) from R&D Systems (Nordenstadt, Germany); the Beta-Glo® Assay System was from Promega (Mannheim, Germany); the β-gal staining kit and cell proliferation reagent WST-1 were obtained from Roche Applied Science (Mannheim, Germany). Vectastain ABC HRP kit (Peroxidase, Standard) and Biotynilated Dolichos Biflorus Agglutinin (DBA) were from Vector Laboratories (Burlingame, CA, USA). The antibody anti-IGF-I (AF-291-NA) was from R&D Systems (Nordenstadt, Germany). Ultra-pure water (resistivity > 18.2 MΩ·cm) was obtained by reverse osmosis (MilliQ®, Millipore Ibérica, Madrid, Spain) and water for cell culture was from Sigma-Aldrich (Steinheim, Germany). All other reagents were analytical grade.
β gal staining kit
Manufactured by Roche
Sourced in Germany
The β-gal staining kit is a laboratory tool used to detect the presence and activity of the β-galactosidase enzyme. It provides a reliable method for visualizing and quantifying β-galactosidase expression in cells or tissues.
Automatically generated - may contain errors
Lab products found in correlation
Beta glo assay system, by Promega (3 mentions)
Aav titration elisa, by Progen Biotechnik (2 mentions)
Cy3 ab labeling kit, by Cytiva (2 mentions)
Biotinylated secondary antibody, by Vector Laboratories (2 mentions)
Cell proliferation reagent wst 1, by Roche (2 mentions)
Anti type x collagen col 10 antibody, by Merck Group (2 mentions)
2 2 azo bis isobutyronitrile, by Thermo Fisher Scientific (1 mentions)
D sucrose, by Thermo Fisher Scientific (1 mentions)
N 3 aminopropyl methacrylamide hydrochloride apma, by Polysciences (1 mentions)
Water for cell culture, by Merck Group (1 mentions)
Dichlorodimethylsilane, by Merck Group (1 mentions)
Anti igf 1 af 291 na, by R&D Systems (1 mentions)
Abc kit, by Vector Laboratories (1 mentions)
Recombinant tgfβ3, by R&D Systems (1 mentions)
Anti sox9 c 20 antibody, by Santa Cruz Biotechnology (1 mentions)
Abc reagent, by Vector Laboratories (1 mentions)
Aavanced concentration reagent, by System Biosciences (1 mentions)
Anti type 1 collagen af 5610 antibody, by OriGene (1 mentions)
4 protocols using β gal staining kit
1
Synthesis and Characterization of Bioactive Hydrogels
2
Chondrogenic Differentiation Assay
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All reagents were from Sigma (Munich, Germany) unless otherwise indicated. Recombinant TGF-β3 was from R&D Systems (Wiesbaden, Germany). The anti-SOX9 (C-20) antibody was from Santa Cruz Biotechnology (Heidelberg, Germany), the anti-type-II collagen (II-II6B3) antibody from the NIH Hybridoma Bank (University of Iowa, Ames, USA), and the anti-type-X collagen (COL-10) antibody from Sigma. Biotinylated secondary antibodies and the ABC kit were from Vector Laboratories (Grünberg, Germany). The β-gal Staining Kit and the Cell Proliferation Reagent WST-1 were from Roche Applied Science (Mannheim, Germany). The Beta-Glo® Assay System was from Promega (Mannheim, Germany). The type-II collagen ELISA (Human COL2A1) was purchased at Cusabio (College Park, MD, USA).
3
Chondrocyte Differentiation Assay
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All reagents were purchased at Sigma (Munich, Germany) unless otherwise indicated. The anti-SOX9 (C-20) and anti-TGF-β (V) antibodies were from Santa Cruz Biotechnology (Heidelberg, Germany), the anti-type-II collagen (II-II6B3) from the NIH Hybridome Bank (University of Iowa, Ames, IA, USA), the anti-type-I collagen (AF-5610) antibody from Acris (Hiddenhausen, Germany), and the anti-type-X collagen (COL-10) antibody from Sigma. The biotinylated secondary antibodies and ABC reagent were from Vector Laboratories (Alexis Deutschland GmbH, Grünberg, Germany). The AAVanced Concentration Reagent was from System Bioscience (Heidelberg, Germany) and the Cy3 Ab Labeling Kit from Amersham/GE Healthcare (Munich, Germany). The AAV titration ELISA was from Progen (Heidelberg, Germany). The β-gal staining kit and the Cell Proliferation Reagent WST-1 were purchased at Roche Applied Science (Mannheim, Germany), the Beta-Glo® Assay System at Promega (Mannheim, Germany), and the TGF-β Quantikine ELISA at R&D Systems (Wiesbaden, Germany).
4
Tumor Tissue Analysis via Histology
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Mice were euthanized at the appropriate endpoint after imaging, considering tumor stage and size. The mice were visually inspected and macroscopically-detectable tumors were collected for further analysis. The skin of the mouse was also compared with the fluorescence image obtained with Maestro, and regions of the skin presenting fluorescence are considered as possible microscopic tumor sites while regions of skin not presenting any fluorescence signal under a certain threshold were considered as possible negative controls.
After removal of tissue, samples were immersed in a 2% formaldehyde, 0.2% glutaraldehyde in PBS solution for at least one hour. The fixative was aspirated and sample washed with PBS twice (PBS aspirated after each washing). The sample was stained using a β-gal staining kit (Roche, cat #: 11828673001) overnight. The staining solution was then removed and the sample washed with a 3% DMSO in PBS solution two times (solution aspirated after each washing), washed with 70% ethanol three times (ethanol aspirated after each washing), and left in 70% ethanol. Samples were processed for paraffin block embedding, sectioning, and hemotoxylin and eosin (H&E) staining (Redwood Dermatopathology, Santa Rosa, CA) [22] (link).
After removal of tissue, samples were immersed in a 2% formaldehyde, 0.2% glutaraldehyde in PBS solution for at least one hour. The fixative was aspirated and sample washed with PBS twice (PBS aspirated after each washing). The sample was stained using a β-gal staining kit (Roche, cat #: 11828673001) overnight. The staining solution was then removed and the sample washed with a 3% DMSO in PBS solution two times (solution aspirated after each washing), washed with 70% ethanol three times (ethanol aspirated after each washing), and left in 70% ethanol. Samples were processed for paraffin block embedding, sectioning, and hemotoxylin and eosin (H&E) staining (Redwood Dermatopathology, Santa Rosa, CA) [22] (link).
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