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e1 plus e2  (Addgene inc)


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    Addgene inc e1 plus e2
    E1 Plus E2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/e1+plus+e2/M24+Nac%3ENac+(Plasmid+%2317174)/10__1128_slash_jvi__02227___16-76-28-57
    Average 92 stars, based on 3 article reviews
    e1 plus e2 - by Bioz Stars, 2026-09
    92/100 stars

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    Polymerase Chain Reaction:

    Article Title: Extracellular Interactions between Hepatitis C Virus and Secreted Apolipoprotein E
    Article Snippet: The DNA fragments were amplified separately and inserted into a pEGFPN2 185 vector (Clontech) using BglII and XbaI to create pN-Env HCV 1–6 plasmids. .. 186 PCR with the primers H77-171-NheF / H77-383-BglIIR, H77-384-NheF/ 187 H77-746-BglIIR, and H77-171-NheF/ H77-746-BglIIR was used to amplify E1 (polyprotein 188 residues 171–383), E2 (polyprotein residues 384–746), and E1 plus E2 (polyprotein residues 189 171–746) from the pN-Env HCV1 template; the products were cleaved by NheI and BglII, 190 ligated with pCW57.1 (a gift from David Root, Addgene plasmid # 41393) and digested by NheI 191 and BamHI to create the lentiviral plasmids pCW-E1, pCW-E2 and pCW-E1E2. .. 192 pEGFP-puro was a gift from Michael McVoy (Addgene plasmid # 45561). pMD2.G and 193 psPAX2 were obtained from Didier Trono (Addgene plasmid # 12259 and 122560).

    Plasmid Preparation:

    Article Title: Extracellular Interactions between Hepatitis C Virus and Secreted Apolipoprotein E
    Article Snippet: The DNA fragments were amplified separately and inserted into a pEGFPN2 185 vector (Clontech) using BglII and XbaI to create pN-Env HCV 1–6 plasmids. .. 186 PCR with the primers H77-171-NheF / H77-383-BglIIR, H77-384-NheF/ 187 H77-746-BglIIR, and H77-171-NheF/ H77-746-BglIIR was used to amplify E1 (polyprotein 188 residues 171–383), E2 (polyprotein residues 384–746), and E1 plus E2 (polyprotein residues 189 171–746) from the pN-Env HCV1 template; the products were cleaved by NheI and BglII, 190 ligated with pCW57.1 (a gift from David Root, Addgene plasmid # 41393) and digested by NheI 191 and BamHI to create the lentiviral plasmids pCW-E1, pCW-E2 and pCW-E1E2. .. 192 pEGFP-puro was a gift from Michael McVoy (Addgene plasmid # 45561). pMD2.G and 193 psPAX2 were obtained from Didier Trono (Addgene plasmid # 12259 and 122560).



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    e1 plus e2  (Addgene inc)
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    Addgene inc e1 plus e2
    E1 Plus E2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/e1+plus+e2/M24+Nac%3ENac+(Plasmid+%2317174)/10__1128_slash_jvi__02227___16-76-28-57
    Average 92 stars, based on 1 article reviews
    e1 plus e2 - by Bioz Stars, 2026-09
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    ccr5 mn e1 e2  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology ccr5 mn e1 e2
    Fig. 1. Models of the <t>CCR5</t> generated by PDBviewer 4.0. (A) CCR5 model showing domains of ECL1, ECL2 and Nt as magenta ribbons and other parts as gray ribbons. (B) CCR5 model showing Nt, ECL1 and incomplete ECL2 contained in CCR5 mN-E1-E2. (C) CCR5 model showing Nt and complete ECL2 contained in N-Linker-E2.
    Ccr5 Mn E1 E2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Fig. 1. Models of the CCR5 generated by PDBviewer 4.0. (A) CCR5 model showing domains of ECL1, ECL2 and Nt as magenta ribbons and other parts as gray ribbons. (B) CCR5 model showing Nt, ECL1 and incomplete ECL2 contained in CCR5 mN-E1-E2. (C) CCR5 model showing Nt and complete ECL2 contained in N-Linker-E2.

    Journal: Protein engineering, design & selection : PEDS

    Article Title: Study on CCR5 analogs and affinity peptides.

    doi: 10.1093/protein/gzr062

    Figure Lengend Snippet: Fig. 1. Models of the CCR5 generated by PDBviewer 4.0. (A) CCR5 model showing domains of ECL1, ECL2 and Nt as magenta ribbons and other parts as gray ribbons. (B) CCR5 model showing Nt, ECL1 and incomplete ECL2 contained in CCR5 mN-E1-E2. (C) CCR5 model showing Nt and complete ECL2 contained in N-Linker-E2.

    Article Snippet: Approximately 500 mg of the recombinant protein CCR5 N-Linker-E2 or CCR5 mN-E1-E2 was added into 40 ml of Protein G PLUS-Agarose (Santa Cruz Biotechnology, USA) and 2 mg of primary antibody CCR5 ECL2 mAb 3A9 (purified mouse IgG2a, k isotype, BD Pharmingen) or CCR5 Nt mAb 2D7 (purified mouse IgG2a, k isotype, BD Pharmingen).

    Techniques: Generated

    Fig. 2. SDS–PAGE (16.5%) of tagged CBD precursors showing expression under various temperature conditions and WB identification. Plasmids pTwin I-CCR5 N-Linker-E2 and pTwin I-CCR5 mN-E1-E2 were transformed into E.coli strains BL21 (DE3). After incubation, the sonicated supernatant and pellet of fusion proteins recovered from equal harvested culture volume were loaded onto each lane. The arrows indicate fusion proteins CCR5 N-Linker-E2-Mxe intein-CBD and CBD-Ssp intein-CCR5 mN-E1-E2 with molecular weights of ≏36.5 and 33.8 kDa, respectively.

    Journal: Protein engineering, design & selection : PEDS

    Article Title: Study on CCR5 analogs and affinity peptides.

    doi: 10.1093/protein/gzr062

    Figure Lengend Snippet: Fig. 2. SDS–PAGE (16.5%) of tagged CBD precursors showing expression under various temperature conditions and WB identification. Plasmids pTwin I-CCR5 N-Linker-E2 and pTwin I-CCR5 mN-E1-E2 were transformed into E.coli strains BL21 (DE3). After incubation, the sonicated supernatant and pellet of fusion proteins recovered from equal harvested culture volume were loaded onto each lane. The arrows indicate fusion proteins CCR5 N-Linker-E2-Mxe intein-CBD and CBD-Ssp intein-CCR5 mN-E1-E2 with molecular weights of ≏36.5 and 33.8 kDa, respectively.

    Article Snippet: Approximately 500 mg of the recombinant protein CCR5 N-Linker-E2 or CCR5 mN-E1-E2 was added into 40 ml of Protein G PLUS-Agarose (Santa Cruz Biotechnology, USA) and 2 mg of primary antibody CCR5 ECL2 mAb 3A9 (purified mouse IgG2a, k isotype, BD Pharmingen) or CCR5 Nt mAb 2D7 (purified mouse IgG2a, k isotype, BD Pharmingen).

    Techniques: SDS Page, Expressing, Transformation Assay, Incubation, Sonication

    Fig. 3. Purification and identification of recombinant proteins CCR5 N-Linker-E2 and CCR5 mN-E1-E2. (A) Intein-mediated recombinant protein CCR5 N-Linker-E2 was purified by CBD chromatography (Peak 1, DTT; Peak 2, CCR5 N-Linker-E2; Peak 3, intein-CBD-tag) followed by Sephadex G-25 to eliminate DTT (Peak 1, DTT; Peak 2, CCR5 N-Linker-E2). SDS–PAGE shows the single band from Peak 2. (B) Intein-mediated purification of the recombinant protein CCR5 mN-E1-E2 by CBD chromatography. SDS–PAGE shows a single band from Peak 1 (Peak 1, mN-E1-E2; Peak 2, CBD-intein-tag). (C) and (D) denote purity and molecular weight of N-Linker-E2 and mN-E1-E2, respectively, determined by HPLC and MS analyses.

    Journal: Protein engineering, design & selection : PEDS

    Article Title: Study on CCR5 analogs and affinity peptides.

    doi: 10.1093/protein/gzr062

    Figure Lengend Snippet: Fig. 3. Purification and identification of recombinant proteins CCR5 N-Linker-E2 and CCR5 mN-E1-E2. (A) Intein-mediated recombinant protein CCR5 N-Linker-E2 was purified by CBD chromatography (Peak 1, DTT; Peak 2, CCR5 N-Linker-E2; Peak 3, intein-CBD-tag) followed by Sephadex G-25 to eliminate DTT (Peak 1, DTT; Peak 2, CCR5 N-Linker-E2). SDS–PAGE shows the single band from Peak 2. (B) Intein-mediated purification of the recombinant protein CCR5 mN-E1-E2 by CBD chromatography. SDS–PAGE shows a single band from Peak 1 (Peak 1, mN-E1-E2; Peak 2, CBD-intein-tag). (C) and (D) denote purity and molecular weight of N-Linker-E2 and mN-E1-E2, respectively, determined by HPLC and MS analyses.

    Article Snippet: Approximately 500 mg of the recombinant protein CCR5 N-Linker-E2 or CCR5 mN-E1-E2 was added into 40 ml of Protein G PLUS-Agarose (Santa Cruz Biotechnology, USA) and 2 mg of primary antibody CCR5 ECL2 mAb 3A9 (purified mouse IgG2a, k isotype, BD Pharmingen) or CCR5 Nt mAb 2D7 (purified mouse IgG2a, k isotype, BD Pharmingen).

    Techniques: Recombinant, Chromatography, SDS Page, Molecular Weight

    Fig. 4. Immunoassay of recombinant extracellular domains of CCR5 analogs. (A) IP of recombinant proteins CCR5 N-Linker-E2 and CCR5 mN-E1-E2 with mAbs 2D7 and 3A9 by silver staining in 16.5% Tris–Tricine–SDS–PAGE. (B) Comparison of exposure epitopes between recombinant proteins CCR5 N-Linker-E2 and CCR5 mN-E1-E2 by indirect ELISA analysis, showing significant differences between the two CCR5 analogs binding to mAb 2D7 and between the two antibodies binding to N-Linker-E2 with P , 0.05. (C) Competitive ELISA of recombinant proteins CCR5 N-Linker-E2 with Met-RANTES. (D) Competitive ELISA of recombinant proteins CCR5 mN-E1-E2 with Met-RANTES.

    Journal: Protein engineering, design & selection : PEDS

    Article Title: Study on CCR5 analogs and affinity peptides.

    doi: 10.1093/protein/gzr062

    Figure Lengend Snippet: Fig. 4. Immunoassay of recombinant extracellular domains of CCR5 analogs. (A) IP of recombinant proteins CCR5 N-Linker-E2 and CCR5 mN-E1-E2 with mAbs 2D7 and 3A9 by silver staining in 16.5% Tris–Tricine–SDS–PAGE. (B) Comparison of exposure epitopes between recombinant proteins CCR5 N-Linker-E2 and CCR5 mN-E1-E2 by indirect ELISA analysis, showing significant differences between the two CCR5 analogs binding to mAb 2D7 and between the two antibodies binding to N-Linker-E2 with P , 0.05. (C) Competitive ELISA of recombinant proteins CCR5 N-Linker-E2 with Met-RANTES. (D) Competitive ELISA of recombinant proteins CCR5 mN-E1-E2 with Met-RANTES.

    Article Snippet: Approximately 500 mg of the recombinant protein CCR5 N-Linker-E2 or CCR5 mN-E1-E2 was added into 40 ml of Protein G PLUS-Agarose (Santa Cruz Biotechnology, USA) and 2 mg of primary antibody CCR5 ECL2 mAb 3A9 (purified mouse IgG2a, k isotype, BD Pharmingen) or CCR5 Nt mAb 2D7 (purified mouse IgG2a, k isotype, BD Pharmingen).

    Techniques: Recombinant, Silver Staining, SDS Page, Comparison, Indirect ELISA, Binding Assay, Competitive ELISA

    Fig. 5. Reactivity between phagotopes isolated by biopanning and the antigen CCR5 N-Linker-E2 or mNE1-E2 determined by capture ELISA (mean and standard deviation of triplicates), showing positive clones (*) with P , 0.05.

    Journal: Protein engineering, design & selection : PEDS

    Article Title: Study on CCR5 analogs and affinity peptides.

    doi: 10.1093/protein/gzr062

    Figure Lengend Snippet: Fig. 5. Reactivity between phagotopes isolated by biopanning and the antigen CCR5 N-Linker-E2 or mNE1-E2 determined by capture ELISA (mean and standard deviation of triplicates), showing positive clones (*) with P , 0.05.

    Article Snippet: Approximately 500 mg of the recombinant protein CCR5 N-Linker-E2 or CCR5 mN-E1-E2 was added into 40 ml of Protein G PLUS-Agarose (Santa Cruz Biotechnology, USA) and 2 mg of primary antibody CCR5 ECL2 mAb 3A9 (purified mouse IgG2a, k isotype, BD Pharmingen) or CCR5 Nt mAb 2D7 (purified mouse IgG2a, k isotype, BD Pharmingen).

    Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Standard Deviation, Clone Assay