Anti-ADAMTS5抗體,軟骨蛋白聚糖抗體免疫組化染色:
電泳(Electrophoresis)
(1)、SDS-PAGE凝膠配制
SDS-PAGE凝膠可以參考一些文獻資料進行配制,也可以使用博研生物生產的SDS-PAGE凝膠配制試劑盒。該試劑盒提供了除水和配膠器具外的所有試劑以及配制各種濃度SDS-PAGE的配方。
(2)、樣品處理
在收集的蛋白樣品中加入適量濃縮的SDS-PAGE蛋白上樣緩沖液。例如2X或5X的SDS-PAGE蛋白上樣緩沖液。使用5X的SDS-PAGE蛋白上樣緩沖液可以減小上樣體積,在相同體積的上樣孔內可以上樣更多的蛋白樣品。5X的SDS-PAGE蛋白上樣緩沖液可以參考相關文獻資料配制,也可以使用我公司生產的SDS-PAGE蛋白上樣緩沖液(5X) 。100℃或沸水浴加熱3-5分鐘,以充分變性蛋白。
(3)、上樣與電泳
冷卻到室溫后,把蛋白樣品直接上樣到SDS-PAGE膠加樣孔內即可。為了便于觀察電泳效果和轉膜效果,以及判斷蛋白分子量大小,使用預染蛋白質分子量標準。
電泳時通常*在上層膠時使用低電壓恒壓電泳,而在溴酚藍進入下層膠時使用高電壓恒壓電泳。對于Bio-Rad的標準電泳裝置或類似電泳裝置,低電壓可以設置在80-100V,高電壓可以設置在120V左右。SDS-PAGE可以采用普通的電泳儀就可以滿足要求。為了電泳方便起見,也可以采用整個SDS-PAGE過程恒壓的方式,通常把電壓設置在100V,然后設定定時時間為90-120分鐘。設置定時可以避免經常發生的電泳過頭。
通常電泳時溴酚藍到達膠的底端處附近即可停止電泳,或者可以根據預染蛋白質分子量標準的電泳情況,預計目的蛋白已經被適當分離后即可停止電泳。
產品名稱:Anti-ADAMTS5抗體,軟骨蛋白聚糖抗體
產品編號:BYK-3573R
英文名稱:Anti-Anti-ADAMTS5
中文名稱:軟骨蛋白聚糖抗體
英文別名:ADAMTS5; A disintegrin and metalloproteinase with thrombospondin motifs 5; A disintegrin like and metalloprotease (reprolysin type) with thrombospondin type 1 motif 5; A Disintigrin And Metalloproteinase with ThromboSpondin motif-5; ADAM metallopeptidase with thrombospondin type 1 motif 5; ADAM TS 11; ADAM TS 5; ADAM TS5; ADAMTS 11; ADAMTS 5; ADAMTS11; ADMP 2; ADMP2; Aggrecanase 2; aggrecanase-2; FLJ36738; Implantin; ThromboSpondin motif-5.
英文介紹:ADAMTS5 was first described as ADAMTS5, a protein elevated in mice during the peri-implantion period. At the same time, another group identified Aggrecanase 11, a protein elevated in arthritic synovium. The name was later changed to ADAMTS5. ADAMTS5 is expressed in human and mouse. It has been found in heart, lung, cervix, uterus, ovary, brain, cartilage, and numerous other tissues, as well as chondroblastoma cell lines. Initial observations indicated a role for ADAMTS5 in aggrecan cleavage and cartilage destruction, especially in arthritis, and potentially a role in embryo implantation. ADAMTS5 contains the canonical HExxHxxxxxH zinc metalloproteinase motif, and has been shown to efficiently cleave Aggrecan. In addition to the metalloprotease domain, ADAMTS5 has a propeptide domain, a prohormone convertase (PC, furin) cleavage site, a cysteine-rich domain, a spacer domain, and two thrombospondin-1 like domains. ADAMTS5 is inhibited by the endogenous MMP inhibitors (TIMP1, 2, 3, and 4) but most efficiently by TIMP3. Unlike many of the ADAMs proteases, ADAMTS5 does not have a transmembrane domain, and is a secreted protein. Full length ADAMTS5 is a 930 amino acid protein with a predicted molecular mass is 101.7 kDa, but glycosylation and the abundance of cysteine residues gives ADAMTS5 a greater apparent molecular weight on reduced SDS PAGE gels. When ADAMTS5 is secreted, it is cleaved at the furin cleavage site (predicted molecular mass 73.2 kDa) and then further cleaved to generate a range of smaller forms.
產品規格:0.1ml/0.2ml
注意事項:This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
此抗體本公司有單抗和多抗,可用于免疫組化(IHC),免疫印跡(WB),酶聯免疫(ELISA),IF、IP、流式細胞術等實驗(可來電索取說明)
此產品本公司有標記的一抗出售,相關標記有:Alexa Fluor 350 標記、Alexa Fluor 488 標記、Alexa Fluor 555 標記、Alexa Fluor 647 標記、AP標記、APC標記、Biotin標記、Cy3標記、Cy5標記、Cy5.5標記、Cy7標記、FITC標記、Gold標記、HRP標記、PE標記、PE-Cy3標記、PE-CY5標記、PE-CY5.5標記、PE-CY7標記、RBITC標記
保存條件:Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.
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