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Zymography is an electrophoretic technique for the detection of hydrolytic enzymes, based on the substrate repertoire of the enzyme. Three types of zymography are used; in gel zymography, in situ zymography and in vivo zymography. [2] For instance, gelatin embedded in a polyacrylamide gel will be digested by active gelatinases run
The next day, the gel is analyzed for regions of gelatin degradation by coomassie blue staining. Patches of gel that are no longer blue following a destain wash are noted. These bands are then correlated to their respective molecular weights in order to identify which cathepsins were active in the sample. Cathepsin K detection by zymography [1]
For electron microscopes, film typically consisted of a gelatin and silver halide emulsion layer on a plastic support base. [2] The silver halide would be converted to silver upon exposure to the electron beam, and the film could then be chemically developed to form an image, which could be digitized for analysis using a film scanner. [2]
Abcam Limited [1] is a producer, distributor and seller of protein research tools operating worldwide from 13 locations with 1,800 employees of which 400 work in Research and Development (R&D). Abcam was listed on the Nasdaq [ 3 ] and the London Stock Exchange until it was acquired by Danaher Corporation in 2023.
Overview of gel electrophoresis. Electrophoresis is a process that enables the sorting of molecules based on charge, size, or shape. Using an electric field, molecules (such as DNA) can be made to move through a gel made of agarose or polyacrylamide.
4313 17390 Ensembl ENSG00000087245 ENSMUSG00000031740 UniProt P08253 P33434 RefSeq (mRNA) NM_004530 NM_001127891 NM_001302508 NM_001302509 NM_001302510 NM_008610 RefSeq (protein) NP_001121363 NP_001289437 NP_001289438 NP_001289439 NP_004521 NP_032636 Location (UCSC) Chr 16: 55.39 – 55.51 Mb Chr 8: 93.55 – 93.58 Mb PubMed search Wikidata View/Edit Human View/Edit Mouse 72 kDa type IV ...
Picture of an SDS-PAGE. The molecular markers (ladder) are in the left lane. Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility.
The substrate in this case is gelatin, and the products are the polypeptides formed. Gelatinase binds to the substrate, gelatin, due to specificity of binding interactions on cell surface. The catalysis, associated with a zinc ion and amino acid residues, breaks the peptide bonds into polypeptides through cleavage.