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產(chǎn)品資料

pcDNA6/V5-His A

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產(chǎn)品名稱: pcDNA6/V5-His A
產(chǎn)品型號(hào):
產(chǎn)品展商: HZbscience
產(chǎn)品文檔: 無(wú)相關(guān)文檔

簡(jiǎn)單介紹

pcDNA6/V5-His A的各批次質(zhì)粒菌株發(fā)貨前均經(jīng)過(guò)嚴(yán)格的多重驗(yàn)證,如存在質(zhì)量問(wèn)題,請(qǐng)?jiān)谑盏疆a(chǎn)品的三個(gè)月內(nèi)通知我司。收到pcDNA6/V5-His A后請(qǐng)短暫離心,取2μl轉(zhuǎn)化至對(duì)應(yīng)感受態(tài)中,挑取單克隆重新提取質(zhì)粒后使用。


pcDNA6/V5-His A  的詳細(xì)介紹

pcDNA6/V5-His A載體基本信息

載體名稱: pcDNA6/V5-His A
質(zhì)粒類型: 哺乳動(dòng)物表達(dá)載體;cDNA表達(dá)載體
高拷貝/低拷貝: 高拷貝
克隆方法: 限制性內(nèi)切酶,多克隆位點(diǎn)
啟動(dòng)子: CMV
載體大小: 5144 bp
5' 測(cè)序引物及序列: T7 Forward: 5’-TAATACGACTCACTATAGGG-3’
3' 測(cè)序引物及序列: BGH Reverse: 5’-TAGAAGGCACAGTCGAGG-3’
載體標(biāo)簽: V5 Epitope(C-端);His tag(C-端)
載體抗性: 氨芐青霉素
篩選標(biāo)記: Blasticidin
克隆菌株: TOP10F′, JM109 and INVaF′
宿主細(xì)胞(系): 常規(guī)細(xì)胞系,如293、Hela等
備注: pcDNA6/V5-His A載體是cDNA的表達(dá)與克隆載體;
CMV啟動(dòng)子驅(qū)動(dòng)目的基因的過(guò)表達(dá);
易于克隆到其它載體中去;
pcDNA6/V5-His A,B,C僅閱讀框不同。
產(chǎn)品目錄號(hào): V220-01
穩(wěn)定性: 瞬表達(dá) 或 穩(wěn)表達(dá)
組成型/誘導(dǎo)型: 組成型
病毒/非病毒: 非病毒

pcDNA6/V5-His A載體質(zhì)粒圖譜和多克隆位點(diǎn)信息

pcDNA6-V5-His A載體圖譜



pcDNA6-V5-His A 多克隆位點(diǎn)

pcDNA6-V5-His 載體特征1
pcDNA6-V5-His 載體特征2

pcDNA6/V5-His A載體簡(jiǎn)介

pcDNA6/V5-His A, B, and C are 5.1 kb vectors derived from pcDNA3.1(+) and designed for high-level expression, selection on blasticidin, purification, and detection of recombinant proteins in mammalian hosts. High-level stable and non-replicative transient expression can be carried out in most mammalian cells. pcDNA6/V5-His A, B, and C 含有以下元件: ? Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a wide range of mammalian cells
? Three reading frames to facilitate in-frame cloning with a C-terminal tag encoding the V5 epitope and a polyhistidine metal-binding peptide
? Blasticidin resistance gene (BSD) from Aspergillus terreus for selection in prokaryotic and eukaryotic cells (Kimura et al., 1994)
? Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen (e.g. COS7)
The control plasmid, pcDNA6/V5-His/lacZ, is the pcDNA6/V5-His A vector with a 3.2 kb fragment containing the β-galactosidase gene cloned in frame with the C-terminal peptide (see page 11). It is included for use as a positive control for transfection, expression, purification, and detection in the cell line of choice. 實(shí)驗(yàn)流程 使用pcDNA.6/V5-His 載體表達(dá)目的基因的實(shí)驗(yàn)流程: 1. Consult the multiple cloning sites described on pages 3–5 to determine which vector (A, B, or C) should be used to clone your gene in frame with the C-terminal V5 epitope and polyhistidine tag.
2. Ligate your insert into the appropriate vector and transform into E. coli. Select transformants on 100 μg/mL blasticidin or 50 to 100 μg/mL ampicillin.
3. Analyze your transformants for the presence of insert by restriction digestion.
4. Select a transformant with the correct restriction pattern and confirm that your gene is in frame with the C-terminal peptide by sequencing.
5. Transfect your construct into the cell line of choice using your own method of transfection.
6. Test for expression of your recombinant gene by western blot analysis or functional assay. If you do not have an antibody to your protein, Life Technologies offers the Anti-V5 Antibody or the Anti-His(C-term) Antibody to detect your recombinant fusion protein.
7. To purify your recombinant fusion protein, you may use metal-chelating resin such as ProBond.. ProBond. resin is available separately. 

pcDNA6/V5-His A載體序列

hz-8629R phospho-Na+/K+-ATPase Alpha1(Ser 943)  磷酸化鈉鉀ATP酶α1多肽抗體
hz-8634R FOXA1/HNF3-alpha  轉(zhuǎn)錄因子HNF-3α抗體
hz-8650R T2R38/TAS2R38  味覺(jué)感受器蛋白T2R38抗體
hz-4156R PRKD1/PKC mu  蛋白激酶C mu型抗體
hz-8654R FAM81A  FAM81A蛋白抗體
hz-5561R phospho-PRKD1(Ser738)  磷酸化蛋白激酶C mu型抗體
hz-8660R Salmonella typhimurium O  傷寒沙門氏菌體蛋白O抗原抗體
hz-8662R BASP1/Nap22  腦富含膜附著信號(hào)蛋白1抗體
hz-8670R Salmonella enteritidis H  傷寒沙門氏菌鞭毛抗原H抗體
hz-5562R phospho-PRKD1(Ser742)  磷酸化蛋白激酶C mu型抗體
hz-8671R MMS19  DNA修復(fù)甲基磺酸敏感性基因19抗體
hz-8672R HDL/high density lipoprotein  高密度脂蛋白抗體
hz-8673R KCTD12  鉀離子通道多聚體結(jié)構(gòu)域蛋白12抗體
hz-8675R BDKRB1  緩激肽B1受體抗體
hz-5563R phospho-PRKD1(Tyr463)  磷酸化蛋白激酶C mu型抗體
hz-8683R IGF2BP1/IMP1  胰島素樣生長(zhǎng)因子ⅡmRNA結(jié)合蛋白1抗體
hz-8686R BRMS1  乳腺癌轉(zhuǎn)移抑制基因1抗體
hz-8687R p53 (FL-393)  腫瘤抑制基因p53抗體
hz-8689R MEPE  細(xì)胞外基質(zhì)磷酸化抗體

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