香焦伊人-香港三级日本三级三级人妇99-香港黄页亚洲一级-香港国产特级一级毛片-闲人吧综合免费888精品-先锋资源亚洲

技術文章您現在的位置:首頁 > 技術文章 > Virogen貨號101-A谷胱甘肽抗體WB檢測文獻數據參考

Virogen貨號101-A谷胱甘肽抗體WB檢測文獻數據參考

更新時間:2025-03-11   點擊次數:84次

谷胱甘肽(glutathione,r-glutamyl cysteingl +glycine,GSH)是一種含γ-酰胺鍵和巰基的三肽,由谷氨酸、半胱氨酸及甘氨酸組成,存在于幾乎身體的每一個細胞。胱甘肽有還原型(GSH)和氧化型(GSSG)兩種形式,在生理條件下以還原型谷胱甘肽占絕大多數。谷胱甘肽還原酶可以催化兩型間的互變。Virogen的谷胱甘肽抗體,貨號101-A,可以檢測谷胱甘肽化蛋白質復合物,當WB檢測時,需要在非還原條件下進行。具體WB檢測的數據,可以參考這篇文獻。

Virogen貨號101-A谷胱甘肽抗體WB檢測文獻數據參考

Antibodies

Glutathionylation was detected with the anti-glutathione antibody MAB5310 (Millipore). The immunoprecipitation of the glutathionylated proteins was carried out with a monoclonal anti-glutathioine (GSH) antibody (#101-A; Virogen). The Na,K-ATPase α1 isoform was immunoprecipitated with SC-21712 antibody (Santa Cruz Biotech., Dallas, TX) and immunedetected with the sc-28800 antibody (H-300; Santa Cruz Biotech.). All α subunits (α(all)) were immunoprecipitated with sc-28800 and immunodetected with the α5 antibody (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City).The β1 isoform was immunodetected with a polyclonal antibody generously provided by Dr. P.A Pedersen, University of Copenhagen. The β2 isoform was detected with the polyclonal antibody 06-1711 (Millipore).


Quantification of glutathionylation

Western blotting of homogenized muscle material has shown that many proteins are susceptible to glutathionylation (Mollica et al. 2012). To study the glutathionylation of Na,K-ATPase subunits it is therefore necessary to use a purification step to isolate the subunits. The level of glutathionylation was studied with two independent techniques.


Method 1

Immunoprecipitation with Na,K-ATPase α and β subunit antibodies. The immunoprecipitate was divided in two parts and used in Western blots both for quantification of α and β subunits with other antibodies and for quantification of glutathionylation with the anti-GSH antibody (MAB5310) (and a sample buffer without the reducing agent dithiothreitol, DTT). The (relative) glutathionylation was calculated as the ratio between the labeling with anti-GSH and the α and β subunit isoform labeling. The glutathionylation of α and β subunit in the homogenate could not be measured with this method due to the presence of other glutathionylated proteins, the yield of glutathionylated α and β subunit proteins could therefore not be calculated.

Method 2

Glutathionylated proteins were immunoprecipitated with the anti-GSH antibody (#101-A; Virogen, Watertown, MA) and afterwards samples were subjected to Western blotting and Na,K-ATPase isoforms were detected with α and β subunit antibodies (sample buffer including DTT). The glutathionylation of α and β subunits was evaluated by calculating the ratio between isoform labeling after immunoblotting (with anti GSH) and the total labeling in the homogenate.



Virogen貨號101-A谷胱甘肽抗體WB檢測文獻數據參考,WB數據圖片:

Virogen貨號101-A谷胱甘肽抗體WB檢測文獻數據參考

The level of α and β subunits glutathionylation, the effects of exercise and terbutaline. The glutathionylated proteins were isolated with immunoprecipitation using anti-glutathione antibodies and Na,K-ATPase subunits quantified with Western blots of the immunoprecipitate (Method 2). (A) H; examples of the β1 subunits detected in the Western blots of two different homogenates (10 μg protein in each lane). IP: β1 subunits detected with anti-β1 antibodies in immunoprecipitates (from100 μg protein) obtained from the same two homogenates using the anti-GSH antibody.



Virogen貨號101-A谷胱甘肽抗體WB檢測文獻數據參考

The relative level of glutathionylated Na,K-ATPase α subunits (Method 1). (A) H; Western blot of muscle homogenate (10 μg protein per lane) labeled with the anti-GSH antibody. IP left: Western blots of two different immunoprecipitates (from 200 μg proteins) using the anti-α(all) antibody (H-300) and labeled on the gel with the anti-α(all) antibody α5. IP right: Western blots of the same two immunoprecipitates labeled with the anti-GSH antibody.





靶點科技(北京)有限公司

靶點科技(北京)有限公司

地址:中關村生命科學園北清創意園2-4樓2層

© 2025 版權所有:靶點科技(北京)有限公司  備案號:京ICP備18027329號-2  總訪問量:286427  站點地圖  技術支持:化工儀器網  管理登陸

主站蜘蛛池模板: 69p69国产精品 | 午夜AV亚洲一码二中文字幕青青 | 国产一区二区三区在线看 | 奇米9999| 暖暖高清日本在线 | 激情亚洲天堂 | 国产福利一区二区在线精品 | 亚洲日本aⅴ片在线观看香蕉 | 成年人免费在线看的惊悚动作片 | 91国产在线第7页 | 国产视频99 | 国内免费高清视频在线观看 | 亚洲精品九色在线网站 | 天天干夜夜添 | 精品91自产拍在线观看99re | 91制片厂制作果冻传媒2021 | 三上悠亚国产精品一区 | 国产亚洲欧美日韩综合综合二区 | 久久伊人在 | 四虎成人免费大片在线 | 日本护士xxxx视频 | 国产欧美曰韩一区二区三区 | 继攵催眠女乱h调教 | 日本videossexx日本人 | 亚洲成人一区 | 亚州在线播放 | 日本888 xxxx| 四虎影院com | aika跟黑人太猛了 | 日本破处 | 国产麻豆剧果冻传媒观看免费视频 | 秘书小说 | jazz中国在线视频 | 亚洲另类第一页 | 国产视频三区 | 国内久久婷婷综合欲色啪 | 男人扒开 | 果冻传媒在线观看的 | 国产精品igao视频网网址 | 深夜精品高中女学生 | 97se亚洲国产综合自在线观看 |