丙烯酰胺

1.Sensitivity of glutathione S-transferases to high doses of acrylamide in albino wistar rats: Affinity purification, biochemical characterization and expression analysis/PMID 31301596; Ecotoxicology and environmental safety 2019 Oct; 182(?):109416/Name matches: sodium dodecyl sulfate acrylamide; polyacrylamide
Abstract:The main objectives of this study were to purify the glutathione S-transfereses (GSTs) and assess the effect of high doses of acrylamide (ACR) on male albino Wistar rat liver, kidney, testis and bran GST activities, and expression analysis of GST. ACR (50 mg/300 ml) was ingested for 40 days (20 doses) in drinking water on alternative days, on 40 day post ingestion the control and treated tissues were collected for GST purification by affinity column and biochemical characterization of GSTs by substrate specificities, and GST expression by immuno dot blots. In the analysis of the purified GSTs, we observed that liver GSTs were resolved in to three bands known as Yc, Yb and Ya; kidney GSTs were resolved in to two bands known as Yc and Ya; testis and brain GSTs were resolved as four bands known as Yc, Yb, Yβ and Yδ on 12.5% sodium dodecyl sulfate polyacrylamide gel (SDS PAGE). In the analysis of biochemical characterization, we observed a significant decrease (p < 0.05) in the specific activities of liver GST isoforms with the substrates 1-chloro 2,4-dinitrobenzene (CDNB), bromosulfophthalein (BSP), p-nitrophenyl acetate (pNPA), p-nitrobenzyl chloride (pNBC) and cumene hydroperoxide (CHP), but showed no activity with ethacrynic acid (ECA) and significant decrease (p < 0.05) in the specific activities of kidney GST isoforms with the substrates CDNB, pNPA, pNBC and CHP, but showed no activity with BSP and ECA, and a significant decrease (p < 0.05) in the specific activities of testis and brain GST isoforms with the substrates CDNB, BSP, pNPA, pNBC, ECA and CHP. In the analysis of immuno dot blots, we observed a decreased expression of liver, kidney, testis and brain GSTs. Through the affinity purification and biochemical characterization, we observed a tissue specific distribution of GSTs that is liver GSTs possess Yc, Yb and Ya sub units known as alpha (α) and mu (μ) class GSTs; kidney GSTs possess Yc and Ya sub units known as (α) alpha class GST; testis and brain GSTs possess Yc, Yb, Yβ and Yδ sub units known as alpha (α), mu (μ) and pi (π) class GSTs. Purification studies, biochemical characterization and immuno dot blot analysis were revealed the GSTs were sensitive to high doses of ACR and the high level exposure to ACR cause the damage of detoxification function of GST due to decreased expression and hence lead to cellular dysfunction of vital organs.

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2.Detecting Protein Antigens in Sodium Dodecyl Sulfate-Polyacrylamide Gels/PMID 31792140; Cold Spring Harbor protocols 2019 12; 2019(12):/Name matches: sodium dodecyl sulfate polyacrylamide
Abstract:In some cases, a native protein can be isolated in its pure form from cell lysates or tissue preparation using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Antigens purified this way often induce good antibody responses. After electrophoresis, the band of protein of interest must be located in the gel. A variety of identification methods can be used, all of which are designed to avoid excessive fixation of the protein in the gel matrix. The choice of method depends partly on the abundance of the polypeptide. Three methods are commonly used: (1) staining side strips cut from the edge of the gel, (2) light staining of the gel itself, and (3) locating the band by radioactive labeling of the antigen. Staining strips of the gel cut from its sides avoids the need to fix the gel. When isolating abundant proteins that are well separated from other bands, staining side strips is a useful method. If the protein is not abundant or is located close to a contaminating band making a clean excision difficult, use one of the other staining methods. If the protein is reasonably abundant, then a light staining of the proteins in the gel with Coomassie Blue G will permit localization without fixing. Alternatively, the bands in the gel can be visualized by immersing the gel in sodium acetate or copper chloride. If the protein is radiolabeled with 125I, 32P, or 35S, then use an autoradiogram as a template to excise the band of interest.

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3.Preparing Protein Antigens from Sodium Dodecyl Sulfate-Polyacrylamide Gels for Immunization/PMID 31792141; Cold Spring Harbor protocols 2019 12; 2019(12):/Name matches: sodium dodecyl sulfate polyacrylamide
Abstract:Some native proteins can be isolated in pure form from cell lysates or tissue preparation using SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Antigens purified this way often induce good antibody responses. There are many different ways to process a gel fragment containing the protein of interest for injection. Such samples can be processed into small pieces and then injected, either by fragmenting the gel by passing it repeatedly through a syringe, or drying the entire gel slice and grinding it into a powder. Injections using the whole gel fragment should only be used with larger animals such as rabbits. For mice or other small animals, electroelution or electrophoretic transfer of the protein should be used to prepare the protein for injection. In the latter method, the protein is transferred to a suitable membrane, such as nitrocellulose or PVDF. The location of the desired protein is identified by staining, and the protein band is then cut from the surrounding membrane, minced, and dissolved in a small amount of dimethyl sulfoxide before subcutaneous injection into the animal.