Ibotenic acid (ibotenate) was purchased from Tocris Bioscience (RD-0285/5, R&D Systems, Germany), isoflurane from Abbvie GmbH (B506100017, Vienna, Austria). LMP (Neurocil®) was obtained from Bayer Pharma (Leverkusen, Germany), cresyl violet acetate from Sigma (C5042, Vienna, Austria). Rabbit polyclonal anti-cleaved caspase-3 antibody was purchased from Cell Signaling Technology (9661, Frankfurt/Main, Germany), biotinylated isolectin B4/G. simplicifolia was obtained from Vector Laboratories (B-1205, Szabo-Scandic, Vienna, Austria), biotinylated goat anti-rabbit IgG from Jackson ImmunoResearch Europe Ltd. (111-065-003 Cambridgeshire, UK), Vectastain Elite ABC Kit from Vector Laboratories (PK-6100 Szabo-Scandic, Vienna, Austria). In situ cell death detection kit, POD (11684817910, TUNEL), recombinant DNase I (04536282001) and proteinase K (03115879001) were obtained from Roche Diagnostics (Mannheim, Germany), diaminobenzidine was purchased from LifeTechnologies (750118, ThermoFisher Scientific, Vienna, Austria), HistoGreen from Linaris (E109, Dossenheim, Germany). Gelatine from cold water fish skin was obtained from Sigma (G7765, Vienna, Austria), biotin-free bovine serum albumin was purchased from Carl Roth (0163.1, Karlsruhe, Germany) and Target Retrieval Solution was from Dako (S1699, Agilent Technologies, Vienna, Austria).
Gelatine from cold water fish skin
Manufactured by Merck Group
Gelatine from cold-water fish skin is a natural, protein-based ingredient derived from the skin of cold-water fish. It is a versatile material with a range of applications in various industries. The core function of this product is to provide a gelling agent, thickener, and stabilizer for a variety of formulations and applications.
Automatically generated - may contain errors
Lab products found in correlation
Albumin from chicken egg white, by Merck Group (2 mentions)
Diaminobenzidine, by Thermo Fisher Scientific (1 mentions)
Cresyl violet acetate, by Merck Group (1 mentions)
Rabbit anti cleaved caspase 3 polyclonal antibody, by Cell Signaling Technology (1 mentions)
Biotin free bovine serum albumin, by Carl Roth (1 mentions)
Target retrieval solution, by Agilent Technologies (1 mentions)
Proteinase k, by Roche (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
Biotinylated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Lr white resin, by Merck Group (1 mentions)
Smt libra 120 plus tem, by Zeiss (1 mentions)
Prolong antifade kit, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by BD (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
4 protocols using gelatine from cold water fish skin
1
Molecular Profiling of Apoptosis in Tissue
2
Ultrastructural Analysis of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The EVs obtained as described above were fixed with Karnovsky’s fixative38 (2.5% (v/v) glutaraldehyde, 2%(v/v) formaldehyde in PBS 1X) 4 h at 4 °C and then dehydrated and embedded in LR White resin (Sigma). The ultrathin section on grids was blocked in a solution containing 1% (w/v) gelatine from cold-water fish skin (SIGMA) prepared in PBS 10 min RT, then, placed on three drops of 0.02 M glycine in PBS for 5 min each of them, and finally blocked with 1% albumin from chicken egg white (Sigma) for 15 min at RT. The grids were placed in a wet chamber and incubated with 20 μl of a 1:20 dilution of anti C-term MASP and anti-clathrin (SCBT) antibodies for 90 min at RT. To eliminate the unbound antibodies, the grids were washed 5× in PBS for 25 min and then immersed in a 1:20 dilution of anti-rat and anti-rabbit polyclonal antibodies labeled with 10 and 25 nm gold-particles, respectively. Afterwards, the grids were washed 4× (5 min each) in PBS followed by washing step of 5 min in distilled water. The samples were finally contrasted with 2% (v/v) uranyl acetate solution and the EM observations were carried out with a Zeiss (Libra- 2000) transmission microscope.
3
Electron Microscopy Imaging of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The pellets with the purified EVs were fixed and treated as previously described19 (link). The grids, containing the ultrathin section of the EVs were blocked in a blocking solution containing 1% (w/v) gelatine from cold-water fish skin (SIGMA) prepared in PBS and incubated for 10 min at RT, then the grids were placed upon of 0.02 M glycine in PBS and finally the blocking procedure concluded with incubation in a solution of 1% albumin from chicken egg white (Sigma) for a total of 15 min at RT.
The grids were placed upon 20 μl for 90 min at RT 1:20 dilution with anti-Clathrin antibodies (SCBT) as EVs putative marker control or anti-MASP SP. The grids were then washed five times in PBS for 25 min. Next, Gold-labelled (10 nm) secondary antibodies were used at 1:20 dilution under the same conditions. Finally, the grids were washed in PBS for 25 min in distilled water and contrasted with 2% (v/v) uranyl acetate solution. EM observations were done in Carl Zeiss SMT LIBRA 120 PLUS TEM.
The grids were placed upon 20 μl for 90 min at RT 1:20 dilution with anti-Clathrin antibodies (SCBT) as EVs putative marker control or anti-MASP SP. The grids were then washed five times in PBS for 25 min. Next, Gold-labelled (10 nm) secondary antibodies were used at 1:20 dilution under the same conditions. Finally, the grids were washed in PBS for 25 min in distilled water and contrasted with 2% (v/v) uranyl acetate solution. EM observations were done in Carl Zeiss SMT LIBRA 120 PLUS TEM.
4
Identifying Adipose-Derived Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To visualise CD34 + /CD90 + cells as indicator for ASCs presence (De Francesco et al., 2009) (link), pure fat and isolated microtissue-SVF were placed into a 12-well plate. Both were washed with PBS, then blocked at room temperature for 2 h with a solution containing PBS, 0.2 % bovine serum albumin (BSA; BD Biosciences) and 0.1 % gelatine from cold-waterfish skin (Sigma-Aldrich). The same solution was used during the incubation with the antibody and for the washing steps, if not indicated differently. The primary antibodies CD34 and CD90 (both 1 µg/ mL; Novus/Bio-Techne Ltd., Abingdon, UK) were applied overnight at 4 °C. On the next day, both conditions were washed and incubated for 4 h at room temperature in the dark with the secondary antibodies goat anti-mouse Rhodamine Red™-X and donkey anti-rabbit Alexa Fluor 488 (both 1 µg/mL; Invitrogen). Nuclear counterstaining was performed with DAPI (0.38 ng/mL; Invitrogen) at room temperature for 10 min in the dark. The samples were washed with PBS and mounted on microscope slides using the Prolong Antifade kit (Molecular Probes). Images were acquired using a confocal microscope (Nikon C2, Nikon).
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!