上海研盟生物科技有限公司Anti-Reprimo抗體*,主要應(yīng)用于WB、IHC、IF、ELISA、流式細(xì)胞術(shù)等實(shí)驗(yàn)中。說明書隨貨發(fā)送,您也可以直接我司在線客服索取??头?/span>
英文名稱:Anti-Reprimo antibody
中文名稱:細(xì)胞質(zhì)內(nèi)的糖基化蛋白抗體
細(xì)胞質(zhì)內(nèi)的糖基化蛋白抗體保存條件: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.
細(xì)胞質(zhì)內(nèi)的糖基化蛋白抗體,Anti-Reprimo抗體抗原修復(fù)方法:
方法1:沸水浴修復(fù),將盛有修復(fù)液和玻片的燒杯置于沸水浴環(huán)境,保持外部沸騰狀態(tài)15min,自然冷卻至室溫。
方法2:微波修復(fù),將盛有修復(fù)液和玻片的燒杯置于微波爐中,高火5min,?;?min,中火5min,自然冷卻至室溫。
方法3:高壓修復(fù),修復(fù)液加入高壓鍋加熱至沸騰,放入玻片,封蓋加壓持續(xù)加熱至噴氣時開始計(jì)時修復(fù)2min,自然冷卻至室溫。
方法4:使用*修復(fù),將*修復(fù)液加入到組織上,37℃,消化30min
★研盟生物★細(xì)胞質(zhì)內(nèi)的糖基化蛋白抗體,Anti-Reprimo抗體
實(shí)驗(yàn)注意事項(xiàng):
1、 把聚丙烯酰胺凝膠中的蛋白質(zhì)電泳轉(zhuǎn)移到硝酸纖維膜上;
a. 轉(zhuǎn)移緩沖液洗滌凝膠和硝酸纖維素膜,將硝酸纖維素膜鋪在凝膠上,用5ml移液管在凝膠上來回滾動去除所有的氣泡
b. 在凝膠/濾膜外再包一張3mm濾紙(預(yù)先用轉(zhuǎn)移緩沖液浸濕),將凝膠夾在中間,保持濕潤和沒有氣泡
c. 將此濾紙/凝膠/薄膜濾紙按照廠家建議方法放入電泳裝置中,凝膠面向陰極
d. 將上述裝置放入緩沖液槽中,并灌滿轉(zhuǎn)移緩沖液以淹沒凝膠
e. 按照廠家所示接通電源開始電泳轉(zhuǎn)移
f. 轉(zhuǎn)移結(jié)束后,取出薄膜和凝膠,棄去凝膠
2、 將薄膜漂在氨基黑中快速染色,直至分子量標(biāo)準(zhǔn)顯現(xiàn)時取出,記錄下標(biāo)準(zhǔn)位置;
3、 用100ml水洗滌纖維素膜,必要時可用脫色緩沖液;
4、 膜置印跡緩沖液中于37℃保溫1小時;
5、 室溫下,用PBS-Tween緩沖液洗滌薄膜
6、 用封口機(jī)將薄膜封入塑料袋中,盡可能不留空氣;
7、 袋的一角剪一緩沖液的小口,用透析袋夾緊;
8、 混合:NGS(100微升),印跡緩沖液中的抗體(2.5-5毫升),加在裝薄膜的袋中,于室溫下?lián)u動2小時(或4℃過ye);
9、 用總體積300ml PBS-Tween緩沖液,分4次在一淺盤中洗滌薄膜,每次75ml;
10、將連接*的羊抗兔IgG(40微升溶于10毫升印跡緩沖液/100微升 NGS)加在袋內(nèi),于室溫下?lián)u動1小時;
11、按步驟9洗滌;
12、加入抗生素蛋白-HRP(40微升溶于10毫升印跡緩沖液/100微升 NGS),于室溫下?lián)u動;
13、Western Blot中轉(zhuǎn)移在膜上的蛋白處于變性狀態(tài),空間結(jié)構(gòu)改變,因此那些識別空間表位的抗體不能用于Western Blot檢測。這種情況可以將表達(dá)目的蛋白的細(xì)胞或細(xì)胞裂解液中的所有蛋白物素化,再用酶標(biāo)記親和素進(jìn)行Western Blot。實(shí)驗(yàn)中取膠和膜需帶手套。
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相關(guān)標(biāo)記抗體:HRP標(biāo)記抗體,Biotin標(biāo)記抗體,Gold標(biāo)記抗體,RBITC標(biāo)記抗體,AP標(biāo)記抗體,FITC標(biāo)記抗體,Cy3標(biāo)記抗體,Cy5標(biāo)記抗體,Cy5.5標(biāo)記抗體,Cy7標(biāo)記抗體,PE標(biāo)記抗體,PE-Cy3標(biāo)記抗體,PE-Cy5標(biāo)記抗體,PE-Cy5.5標(biāo)記抗體,PE-Cy7標(biāo)記抗體,APC標(biāo)記抗體,Alexa Fluor 350標(biāo)記抗體,Alexa Fluor 488標(biāo)記抗體,Alexa Fluor 555標(biāo)記抗體,Alexa Fluor 647標(biāo)記抗體
簡單介紹:
產(chǎn)品別名:Protein reprimo.; reprimo, TP53 dependent G2 arrest mediator candidate; 2410012A13RIK; FLJ90327; MGC109515; REPRIMO; RPRM; RPRM_HUMAN.
抗體來源:Rabbit or Mouse
保質(zhì)期:1年
克隆類型:Polyclonal or monoclonal
性 狀:Lyophilized or Liquid
濃 度:1mg/1ml
背景介紹:Reprimo, also known as TP53 dependent G2 arrest mediator candidate or RPRM, is a 109 amino acid cytoplasmic protein belonging to the Reprimo family, which is involved in the regulation of p53-dependent G2 arrest of the cell cycle. Reprimo may initiate cell cycle arrest by inhibiting CDC2 and nuclear translocation of the CDC2 cyclin B1 complex. A highly glycosylated protein, Reprimo also plays a role in cell cycle surveillance and DNA repair. Hypermethylation of Reprimo can lead to its transcriptional repression, which may increase pathogenesis of some types of human cancers. Reprimo has been identified as a potential biomarker for early detection multiple cancers. The gene encoding Reprimo maps to human chromosome 2, which houses over 1,400 genes and comprises nearly 8% of the human genome.
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