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SignaGen lipid transfection
Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
Lipid Transfection, supplied by SignaGen, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Nef-mediated enhancement of cellular activation and human immunodeficiency virus type 1 replication in primary T cells is dependent on association with p21-activated kinase 2

Journal: Retrovirology

doi: 10.1186/1742-4690-8-64

Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h post-transfection, cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
Figure Legend Snippet: Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h post-transfection, cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.

Techniques Used: Activation Assay, Transfection, Plasmid Preparation, Lysis, Expressing, Western Blot

Related Articles

Transfection:

Article Title: Nef-mediated enhancement of cellular activation and human immunodeficiency virus type 1 replication in primary T cells is dependent on association with p21-activated kinase 2
Article Snippet: .. 3.5 μg of pHAGE vector,3.5 μg of the packaging construct pDR89.1 [ ], and 1 μg of pVSV-G were transfected by lipid transfection (LipoD293T, Signagen) into 5.5 × 106 293T cells plated 24 h prior in a 10-cm tissue culture plate. ..

Plasmid Preparation:

Article Title: Nef-mediated enhancement of cellular activation and human immunodeficiency virus type 1 replication in primary T cells is dependent on association with p21-activated kinase 2
Article Snippet: .. 3.5 μg of pHAGE vector,3.5 μg of the packaging construct pDR89.1 [ ], and 1 μg of pVSV-G were transfected by lipid transfection (LipoD293T, Signagen) into 5.5 × 106 293T cells plated 24 h prior in a 10-cm tissue culture plate. ..

Construct:

Article Title: Nef-mediated enhancement of cellular activation and human immunodeficiency virus type 1 replication in primary T cells is dependent on association with p21-activated kinase 2
Article Snippet: .. 3.5 μg of pHAGE vector,3.5 μg of the packaging construct pDR89.1 [ ], and 1 μg of pVSV-G were transfected by lipid transfection (LipoD293T, Signagen) into 5.5 × 106 293T cells plated 24 h prior in a 10-cm tissue culture plate. ..

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  • 88
    SignaGen lipid transfection
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Lipid Transfection, supplied by SignaGen, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipid transfection/product/SignaGen
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    lipid transfection - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    85
    SignaGen genjet lipid transfection reagents
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Genjet Lipid Transfection Reagents, supplied by SignaGen, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/genjet lipid transfection reagents/product/SignaGen
    Average 85 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    genjet lipid transfection reagents - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

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    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h post-transfection, cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.

    Journal: Retrovirology

    Article Title: Nef-mediated enhancement of cellular activation and human immunodeficiency virus type 1 replication in primary T cells is dependent on association with p21-activated kinase 2

    doi: 10.1186/1742-4690-8-64

    Figure Lengend Snippet: Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h post-transfection, cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.

    Article Snippet: 3.5 μg of pHAGE vector,3.5 μg of the packaging construct pDR89.1 [ ], and 1 μg of pVSV-G were transfected by lipid transfection (LipoD293T, Signagen) into 5.5 × 106 293T cells plated 24 h prior in a 10-cm tissue culture plate.

    Techniques: Activation Assay, Transfection, Plasmid Preparation, Lysis, Expressing, Western Blot