上海研盟生物科技有限公司Anti-TERT抗體,端粒酶逆轉錄酶抗體*,主要應用于WB、IHC、IF、ELISA、流式細胞術等實驗中。說明書隨貨發送,您也可以直接我司在線客服索取??头?/p>
簡單介紹:
中文名稱:端粒酶逆轉錄酶抗體
英文名稱:Anti-TERT
產品編號:byk-1411R
產品別名:EST2; hEST2; TCS1; omerase associated protein 2; omerase Catalytic Subunit; omere Reverse Transcriptase; omerase reverse transcriptase; TERT; TP2; TRT; TERT_HUMAN; omerase reverse transcriptase; HEST2; omerase catalytic subunit; omerase-associated protein 2.
產品用途:科研實驗
產品價格:請報價
說明書:請添加客服直接索取
抗體來源:該指標有兩種產品,一種是兔來源抗體,一種是鼠來源抗體
克隆類型:兔來源為多克隆抗體,鼠來源單克隆抗體
交叉反應:請索取說明書查看
性 狀:Lyophilized or Liquid
濃 度:1mg/1ml
亞 型:IgG
純化方法: affinity purified by Protein A
儲 存 液: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol
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.
Anti-TERT抗體,端粒酶逆轉錄酶抗體產品介紹:omerase is a ribonucleoprotein enzyme essential for the replication of chromosome termini in most eukaryotes. It elongates omeres. It is a reverse transcriptase that adds simple sequence repeats to chromosome ends by copying a template sequence within the RNA component of the enzyme. omerase are large DNA-protein complexes with omerase expression being the subject of recent research due to its link to cell immortalization. Recent evidence has shown that MYC upregulates the catalytic subunit of omerase, TERT, and that omerase is a ribonucleoprotein polymerase that maintains omere ends by addition of the omere repeat TTAGGG. The enzyme consists of a protein component with reverse transcriptase activity, encoded by this gene, and an RNA component which serves as a template for the omere repeat. omerase expression plays a role in cellular senescence, as it is normally repressed in postnatal somatic cells resulting in progressive shortening of omeres. Deregulation of omerase expression in somatic cells may be involved in oncogenesis. Studies in mouse suggest that omerase also participates in chromosomal repair, since de novo synthesis of omere repeats may occur at double-stranded breaks. Alternatively spliced variants encoding different isoforms of omerase reverse transcriptase have been identified; the full-length sequence of some variants has not been determined. Alternative splicing at this locus is thought to be one mechanism of regulation of omerase activity.
Western-blotting試驗方法:
蛋白提樣品制備
1、 將培養液吸入至15 ml離心管中,于2500 rpm離心5 min。
2、 棄上清,加入5 ml PBS并用槍輕輕吹打洗滌,然后2500 rpm離心5 min。棄上清后用PBS重復洗滌一次。
3、 用槍吸干上清后,加100 μl裂解液(含PMSF)冰上裂解30 min,裂解過程中要經常彈一彈以使細胞充分裂解。
4、將裂解液4℃、12000 rpm離心5 min,取上清分裝于0.5 ml EP管中并置于-20℃保存。
5、BCA法測蛋白濃度
SDS-PAGE電泳
1、 玻璃板對齊后放入夾中卡緊,然后垂直卡在架子上準備灌膠。
2、按前面方法配10%分離膠,加入TEMED后立即搖勻即可灌膠。灌膠時,可用10 ml槍吸取5 ml膠沿玻璃放出,待膠面升到綠帶中間線高度時即可。然后膠上加一層水,液封后的膠凝的更快。(灌膠時開始可快一些,膠面快到所需高度時要放慢速度。操作時膠一定要沿玻璃板流下,這樣膠中才不會有氣泡。加水液封時要慢,否則膠會被沖變型。)
3、 當水和膠之間有一條折射線時,說明膠已凝了。再等3 min使膠充分凝固就可倒去膠上層水并用吸水紙將水吸干。
4、 按方法配4%的濃縮膠,加入TEMED后立即搖勻即可灌膠。將剩余空間灌滿濃縮膠后將梳子插入孔槽。灌膠時也要使膠沿玻璃板流下以免膠中有氣泡產生。插梳子時要使梳子保持水平。待到濃縮膠凝固后,兩手分別捏住梳子的兩邊豎直向上輕輕將其拔出。
5、 用水沖洗濃縮膠,將其放入電泳槽中。
6、 測完蛋白含量后,計算含20-50 μg蛋白(根據自己的實驗需要進行選擇,沒有固定的量)的溶液體積即為上樣量。取出上樣樣品至200 μl的EP管中,加入5×SDS 上樣緩沖液至終濃度為1×。上樣前要將樣品于沸水中煮5-10 min使蛋白變性。
7、 加入足夠的電泳緩沖液后開始準備上樣。(電泳液至少要漫過內測的小玻璃板。)用微量進樣器貼壁吸取樣品,將樣品吸出不要吸進氣泡。將加樣器針頭插至加樣孔中緩慢加入樣品(包括一孔做可視的蛋白marker)。(加樣太快可使樣品沖出加樣孔,若有氣泡也可能使樣品溢出。加入下一個樣品時,進樣器需在外槽電泳緩沖液中洗滌數次,以免交叉污染)
8、電泳:濃縮膠時用80-100 V跑,到分離膠以后用150-200 V跑,總時間2 h左右,電泳至溴酚蘭剛跑出即可終止電泳,進行轉膜。
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