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pcDNA4/myc-His A

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產品名稱: pcDNA4/myc-His A
產品型號:
產品展商: HZbscience
產品文檔: 無相關文檔

簡單介紹

pcDNA4/myc-His A的各批次質粒菌株發貨前均經過嚴格的多重驗證,如存在質量問題,請在收到產品的三個月內通知我司。收到pcDNA4/myc-His A后請短暫離心,取2μl轉化至對應感受態中,挑取單克隆重新提取質粒后使用。


pcDNA4/myc-His A  的詳細介紹

pcDNA4/myc-His A載體基本信息

載體名稱: pcDNA4/myc-His A
質粒類型: 哺乳動物表達載體;cDNA表達載體
高拷貝/低拷貝: 高拷貝
克隆方法: 多克隆位點,限制性內切酶
啟動子: CMV
載體大小: 5075 bp
5' 測序引物及序列: T7 Forward: 5’-TAATACGACTCACTATAGGG-3’
3' 測序引物及序列: BGH Reverse: 5-TAGAAGGCACAGTCGAGG-3
載體標簽: His Tag (C-端), c-Myc Epitope Tag(C-端)
載體抗性: 氨芐青霉素
篩選標記: Zeocin
克隆菌株: TOP10F′, DH5a, JM109, TOP10
宿主細胞(系): 常規細胞系,如293、Hela等
備注: pcDNA4/myc-His A 載體是哺乳動物表達載體,適用于cDNA的表達與克隆;
CMV啟動子驅動目的基因的高水平表達;
pcDNA4/myc-His A,B,C的 區別僅在于多克隆位點處;
產品目錄號: V863-20
穩定性: 瞬表達 或 穩表達
組成型/誘導型: 組成型
病毒/非病毒: 非病毒

pcDNA4/myc-His A載體質粒圖譜和多克隆位點信息

pcDNA4-myc-His A載體圖譜



pcDNA4-myc-His A 多克隆位點

pcDNA4-myc-His 載體特征

pcDNA4/myc-His A載體簡介

載體描述

pcDNA4/myc-His A, B, and C are 5.1 kb vectors designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins (see pages 11-12 for more information). High-level stable and transient expression can be carried out in most mammalian cells. The vectors contain the following elements:
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 peptide encoding the myc (c-myc) epitope and a polyhistidine (6xHis) metal-binding tag
Zeocin resistance gene for selection of stable cell lines (Mulsant et al., 1988) (see page 14 for more information).
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, pcDNA4/myc-His/lacZ is included for use as a positive control for transfection, expression, and detection in the cell line of choice.

實驗流程:

Use the following outline to clone and express your gene of interest in pcDNA4/myc-His:
1.Consult the multiple cloning sites described on pages 3-4 to determine which vector (A, B, or C) to use for cloning your gene in frame with the C-terminal myc epitope and the polyhistidine tag.
2.Ligate your insert into the appropriate vector and transform into E. coli. Select transformants on 50 to 100 μg/mL ampicillin or 25 to 50g/mL Zeocin in Low Salt LB. For more information.
3.Analyze your transformants for the presence of insert by restriction digestion.
4.Select a transformant with the correct restriction pattern and use sequencing to confirm that your gene is cloned in-frame with the C-terminal peptide.
5.Transfect your construct into the cell line of choice using your own method of transfection. Generate a stable cell line, if desired.
6.Test for expression of your recombinant gene by western blot analysis or functional assay. For antibodies to the myc epitope or the C-terminal polyhistidine tag.
7.To purify your recombinant protein, you may use metal-chelating resin such as ProBond. ProBond resin is available separately 

pcDNA4/myc-His A載體序列

hz-9366R PSMD7  蛋白酶調解因子7抗體
hz-9367R PSMD6/P44S10  蛋白酶調解因子6抗體
hz-9368R OCEL1  緊密連接蛋白/小分子膠原蛋白OCEL1抗體
hz-9369R ODF3B  外致密纖維蛋白3B抗體
hz-9370R PGBD1/Cerebral protein 4  腦蛋白4抗體
hz-9371R PGBD3  PGBD3蛋白抗體
hz-9372R PGGT1β  香葉烯基轉移酶1β抗體
hz-9375R MYCBP2  MYC結合蛋白2抗體
hz-9376R MURF3  肌肉特定環指蛋白3抗體
hz-9377R MMS2/EDPF1  腸細胞分化相關因子1抗體
hz-9378R MKRN1/RNF61  環指蛋白61抗體
hz-9379R MKRN2/RNF62  環指蛋白62抗體
hz-9380R MID1/Midline-1/RNF59  環指蛋白59抗體
hz-9382R KF1/ZFP103  鋅指蛋白103抗體
hz-9383R CNOT2  細胞表面趨化因子受體4相關蛋白2抗體
hz-9384R phospho-CNOT2(Ser101)  磷酸化細胞表面趨化因子受體4相關蛋白2抗體
hz-9285R CNOT4  細胞表面趨化因子受體4相關蛋白4抗體
hz-9386R CNOT6  細胞表面趨化因子受體4相關蛋白6抗體
hz-9387R LRSAM1  LRSAM1蛋白抗體
hz-9388R CNOT8  細胞表面趨化因子受體4相關蛋白8抗體
hz-9389R MURF2  肌肉特異性環指蛋白2抗體
hz-9390R MURF3  肌肉特異性環指蛋白3抗體

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