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egfp sec23a  (Addgene inc)


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    Structured Review

    Addgene inc egfp sec23a
    (A) Proximity labeling of HEK293 ATG2 DKO cells stably expressing APEX2-GFP-ATG2A was performed as previously described . Three biological replicates were averaged and plotted as the log2 ratio between the full labeling reaction in complete media over cells that were not treated with hydrogen peroxide on the x axis, and -log10 of the p-value on the y axis. The red line demonstrates the p-value cutoff of 0.05. The proteomics revealed an enrichment of autophagy (labeled in green) and early secretory proteins (COPII/ER Exit Site proteins are labeled in orange, ARFGAP1 is labeled in red). (B) Strategy to identify phagophores by live cell imaging. Phagophores are defined as sites where both GFP-ATG2A and TagBFP-LC3B co-localize. (C-D) Live cell imaging of ATG2 DKO cells stably expressing GFP-ATG2A and TagBFP-LC3B and transfected with <t>mRuby-Sec23A</t> or RFP-Sec23IP/p125a or ARFGAP1-TagRFP. The cells were starved for 2-4 hours prior to imaging. The fraction of phagophores (white arrows) that colocalized with the third fluorescent protein (transfected gene) was quantified and plotted in (D). Statistical significance was assessed by one way ANOVA. *, adjusted P value <0.05. **, adjusted P value <0.01. ***, adjusted P value <0.001. ****, adjusted P value <0.0001. Data from three biological replicates were pooled for each condition. Maximum intensity projections of confocal images. (E) Global organelle profiling data from https://organelles.sf.czbiohub.org . Whole organelle profiles were established by pulling down endogenously tagged proteins and performing quantitative proteomics. The enrichment value for each protein were calculated by taking the difference in median log2 LFQ values between the replicates and the null distribution.
    Egfp Sec23a, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egfp sec23a/product/Addgene inc
    Average 93 stars, based on 15 article reviews
    egfp sec23a - by Bioz Stars, 2026-03
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    Images

    1) Product Images from "ATG2A engages Rab1a and ARFGAP1 positive membranes during autophagosome biogenesis"

    Article Title: ATG2A engages Rab1a and ARFGAP1 positive membranes during autophagosome biogenesis

    Journal: bioRxiv

    doi: 10.1101/2025.03.24.645038

    (A) Proximity labeling of HEK293 ATG2 DKO cells stably expressing APEX2-GFP-ATG2A was performed as previously described . Three biological replicates were averaged and plotted as the log2 ratio between the full labeling reaction in complete media over cells that were not treated with hydrogen peroxide on the x axis, and -log10 of the p-value on the y axis. The red line demonstrates the p-value cutoff of 0.05. The proteomics revealed an enrichment of autophagy (labeled in green) and early secretory proteins (COPII/ER Exit Site proteins are labeled in orange, ARFGAP1 is labeled in red). (B) Strategy to identify phagophores by live cell imaging. Phagophores are defined as sites where both GFP-ATG2A and TagBFP-LC3B co-localize. (C-D) Live cell imaging of ATG2 DKO cells stably expressing GFP-ATG2A and TagBFP-LC3B and transfected with mRuby-Sec23A or RFP-Sec23IP/p125a or ARFGAP1-TagRFP. The cells were starved for 2-4 hours prior to imaging. The fraction of phagophores (white arrows) that colocalized with the third fluorescent protein (transfected gene) was quantified and plotted in (D). Statistical significance was assessed by one way ANOVA. *, adjusted P value <0.05. **, adjusted P value <0.01. ***, adjusted P value <0.001. ****, adjusted P value <0.0001. Data from three biological replicates were pooled for each condition. Maximum intensity projections of confocal images. (E) Global organelle profiling data from https://organelles.sf.czbiohub.org . Whole organelle profiles were established by pulling down endogenously tagged proteins and performing quantitative proteomics. The enrichment value for each protein were calculated by taking the difference in median log2 LFQ values between the replicates and the null distribution.
    Figure Legend Snippet: (A) Proximity labeling of HEK293 ATG2 DKO cells stably expressing APEX2-GFP-ATG2A was performed as previously described . Three biological replicates were averaged and plotted as the log2 ratio between the full labeling reaction in complete media over cells that were not treated with hydrogen peroxide on the x axis, and -log10 of the p-value on the y axis. The red line demonstrates the p-value cutoff of 0.05. The proteomics revealed an enrichment of autophagy (labeled in green) and early secretory proteins (COPII/ER Exit Site proteins are labeled in orange, ARFGAP1 is labeled in red). (B) Strategy to identify phagophores by live cell imaging. Phagophores are defined as sites where both GFP-ATG2A and TagBFP-LC3B co-localize. (C-D) Live cell imaging of ATG2 DKO cells stably expressing GFP-ATG2A and TagBFP-LC3B and transfected with mRuby-Sec23A or RFP-Sec23IP/p125a or ARFGAP1-TagRFP. The cells were starved for 2-4 hours prior to imaging. The fraction of phagophores (white arrows) that colocalized with the third fluorescent protein (transfected gene) was quantified and plotted in (D). Statistical significance was assessed by one way ANOVA. *, adjusted P value <0.05. **, adjusted P value <0.01. ***, adjusted P value <0.001. ****, adjusted P value <0.0001. Data from three biological replicates were pooled for each condition. Maximum intensity projections of confocal images. (E) Global organelle profiling data from https://organelles.sf.czbiohub.org . Whole organelle profiles were established by pulling down endogenously tagged proteins and performing quantitative proteomics. The enrichment value for each protein were calculated by taking the difference in median log2 LFQ values between the replicates and the null distribution.

    Techniques Used: Labeling, Stable Transfection, Expressing, Live Cell Imaging, Transfection, Imaging



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    (A) Proximity labeling of HEK293 ATG2 DKO cells stably expressing APEX2-GFP-ATG2A was performed as previously described . Three biological replicates were averaged and plotted as the log2 ratio between the full labeling reaction in complete media over cells that were not treated with hydrogen peroxide on the x axis, and -log10 of the p-value on the y axis. The red line demonstrates the p-value cutoff of 0.05. The proteomics revealed an enrichment of autophagy (labeled in green) and early secretory proteins (COPII/ER Exit Site proteins are labeled in orange, ARFGAP1 is labeled in red). (B) Strategy to identify phagophores by live cell imaging. Phagophores are defined as sites where both GFP-ATG2A and TagBFP-LC3B co-localize. (C-D) Live cell imaging of ATG2 DKO cells stably expressing GFP-ATG2A and TagBFP-LC3B and transfected with <t>mRuby-Sec23A</t> or RFP-Sec23IP/p125a or ARFGAP1-TagRFP. The cells were starved for 2-4 hours prior to imaging. The fraction of phagophores (white arrows) that colocalized with the third fluorescent protein (transfected gene) was quantified and plotted in (D). Statistical significance was assessed by one way ANOVA. *, adjusted P value <0.05. **, adjusted P value <0.01. ***, adjusted P value <0.001. ****, adjusted P value <0.0001. Data from three biological replicates were pooled for each condition. Maximum intensity projections of confocal images. (E) Global organelle profiling data from https://organelles.sf.czbiohub.org . Whole organelle profiles were established by pulling down endogenously tagged proteins and performing quantitative proteomics. The enrichment value for each protein were calculated by taking the difference in median log2 LFQ values between the replicates and the null distribution.
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    pegfp sec23a  (Addgene inc)
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    (A) Proximity labeling of HEK293 ATG2 DKO cells stably expressing APEX2-GFP-ATG2A was performed as previously described . Three biological replicates were averaged and plotted as the log2 ratio between the full labeling reaction in complete media over cells that were not treated with hydrogen peroxide on the x axis, and -log10 of the p-value on the y axis. The red line demonstrates the p-value cutoff of 0.05. The proteomics revealed an enrichment of autophagy (labeled in green) and early secretory proteins (COPII/ER Exit Site proteins are labeled in orange, ARFGAP1 is labeled in red). (B) Strategy to identify phagophores by live cell imaging. Phagophores are defined as sites where both GFP-ATG2A and TagBFP-LC3B co-localize. (C-D) Live cell imaging of ATG2 DKO cells stably expressing GFP-ATG2A and TagBFP-LC3B and transfected with <t>mRuby-Sec23A</t> or RFP-Sec23IP/p125a or ARFGAP1-TagRFP. The cells were starved for 2-4 hours prior to imaging. The fraction of phagophores (white arrows) that colocalized with the third fluorescent protein (transfected gene) was quantified and plotted in (D). Statistical significance was assessed by one way ANOVA. *, adjusted P value <0.05. **, adjusted P value <0.01. ***, adjusted P value <0.001. ****, adjusted P value <0.0001. Data from three biological replicates were pooled for each condition. Maximum intensity projections of confocal images. (E) Global organelle profiling data from https://organelles.sf.czbiohub.org . Whole organelle profiles were established by pulling down endogenously tagged proteins and performing quantitative proteomics. The enrichment value for each protein were calculated by taking the difference in median log2 LFQ values between the replicates and the null distribution.
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    sec23a plasmid  (Addgene inc)
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    ( A ) Volcano plot of the TMED9 interactome (Myc-TMED9 + MUC1-fs) shows preferential binding of TMED9 (green dot) to COPI proteins (red dots), but not to COPII proteins (purple dots), which did not pass the significance threshold for enrichment compared to the negative control (empty vector + MUC1-fs). ( B ) Myc-tagged TMED9 co-IP’ed. COPB2 (COPI protein) but not SEC13 (COPII protein). β-Actin is shown as a loading control in the lysate and as a negative control in the pulldown. Blots are representative of three independent experiments. ( C ) Myc-tagged TMED9 mutant R223E coimmunoprecipitated less endogenous COPB2 than WT TMED9. ( D ) GFP-tagged <t>SEC23a</t> coimmunoprecipitated mutant and WT TMED9. ** P < 0.05. ns., not significant.
    Sec23a Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pegfpsec23a  (Addgene inc)
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    Addgene inc pegfpsec23a
    ( A ) Volcano plot of the TMED9 interactome (Myc-TMED9 + MUC1-fs) shows preferential binding of TMED9 (green dot) to COPI proteins (red dots), but not to COPII proteins (purple dots), which did not pass the significance threshold for enrichment compared to the negative control (empty vector + MUC1-fs). ( B ) Myc-tagged TMED9 co-IP’ed. COPB2 (COPI protein) but not SEC13 (COPII protein). β-Actin is shown as a loading control in the lysate and as a negative control in the pulldown. Blots are representative of three independent experiments. ( C ) Myc-tagged TMED9 mutant R223E coimmunoprecipitated less endogenous COPB2 than WT TMED9. ( D ) GFP-tagged <t>SEC23a</t> coimmunoprecipitated mutant and WT TMED9. ** P < 0.05. ns., not significant.
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    Image Search Results


    (A) Proximity labeling of HEK293 ATG2 DKO cells stably expressing APEX2-GFP-ATG2A was performed as previously described . Three biological replicates were averaged and plotted as the log2 ratio between the full labeling reaction in complete media over cells that were not treated with hydrogen peroxide on the x axis, and -log10 of the p-value on the y axis. The red line demonstrates the p-value cutoff of 0.05. The proteomics revealed an enrichment of autophagy (labeled in green) and early secretory proteins (COPII/ER Exit Site proteins are labeled in orange, ARFGAP1 is labeled in red). (B) Strategy to identify phagophores by live cell imaging. Phagophores are defined as sites where both GFP-ATG2A and TagBFP-LC3B co-localize. (C-D) Live cell imaging of ATG2 DKO cells stably expressing GFP-ATG2A and TagBFP-LC3B and transfected with mRuby-Sec23A or RFP-Sec23IP/p125a or ARFGAP1-TagRFP. The cells were starved for 2-4 hours prior to imaging. The fraction of phagophores (white arrows) that colocalized with the third fluorescent protein (transfected gene) was quantified and plotted in (D). Statistical significance was assessed by one way ANOVA. *, adjusted P value <0.05. **, adjusted P value <0.01. ***, adjusted P value <0.001. ****, adjusted P value <0.0001. Data from three biological replicates were pooled for each condition. Maximum intensity projections of confocal images. (E) Global organelle profiling data from https://organelles.sf.czbiohub.org . Whole organelle profiles were established by pulling down endogenously tagged proteins and performing quantitative proteomics. The enrichment value for each protein were calculated by taking the difference in median log2 LFQ values between the replicates and the null distribution.

    Journal: bioRxiv

    Article Title: ATG2A engages Rab1a and ARFGAP1 positive membranes during autophagosome biogenesis

    doi: 10.1101/2025.03.24.645038

    Figure Lengend Snippet: (A) Proximity labeling of HEK293 ATG2 DKO cells stably expressing APEX2-GFP-ATG2A was performed as previously described . Three biological replicates were averaged and plotted as the log2 ratio between the full labeling reaction in complete media over cells that were not treated with hydrogen peroxide on the x axis, and -log10 of the p-value on the y axis. The red line demonstrates the p-value cutoff of 0.05. The proteomics revealed an enrichment of autophagy (labeled in green) and early secretory proteins (COPII/ER Exit Site proteins are labeled in orange, ARFGAP1 is labeled in red). (B) Strategy to identify phagophores by live cell imaging. Phagophores are defined as sites where both GFP-ATG2A and TagBFP-LC3B co-localize. (C-D) Live cell imaging of ATG2 DKO cells stably expressing GFP-ATG2A and TagBFP-LC3B and transfected with mRuby-Sec23A or RFP-Sec23IP/p125a or ARFGAP1-TagRFP. The cells were starved for 2-4 hours prior to imaging. The fraction of phagophores (white arrows) that colocalized with the third fluorescent protein (transfected gene) was quantified and plotted in (D). Statistical significance was assessed by one way ANOVA. *, adjusted P value <0.05. **, adjusted P value <0.01. ***, adjusted P value <0.001. ****, adjusted P value <0.0001. Data from three biological replicates were pooled for each condition. Maximum intensity projections of confocal images. (E) Global organelle profiling data from https://organelles.sf.czbiohub.org . Whole organelle profiles were established by pulling down endogenously tagged proteins and performing quantitative proteomics. The enrichment value for each protein were calculated by taking the difference in median log2 LFQ values between the replicates and the null distribution.

    Article Snippet: The following plasmids were used in this study: pLVX Puro GFP-ATG2A (Previously published ), pLVX Puro APEX2-GFP-ATG2A (This study; a PCR fragment containing flanking EcoRI cutsites around APEX2 was generated from pcDNA3 APEX2-NES (Addgene #49386) and was inserted into GFP-ATG2A by ligation), pCMV 3xFLAG-ATG2A (Previously published ), pLVX Neo TagBFP-LC3B (This study; pLVX puro TagBFP-LC3B and pLVX-EGFP-IRES-Neo (Addgene #128660) were cut with EcoRI and BamHI, following which the TagBFP-LC3B fragment was ligated into the pLVX Neo backbone), pDESt47 Sar1-GFP (Addgene #67409), Sar1b-mOrange2 (Addgene #166899), Sar1bH79G2-mOrange2 (Addgene #166900), Sar1bH79G-3xFLAG (This Study; A BamHI-3xFLAG-NotI fragment was generated by oligo annealing, following which it was ligated into the Sar1bH79G-mOrange2 backbone that was cut by the same restriction enzymes), mRuby-Sec23A (Addgene 36158), EGFP-Sec23A (Addgene #66609), pEGFP-Sec24A (This Study; a XhoI-Sec24A-PacI fragment was generated by PCR from HA-Sec24A which was a generous gift from the Kundu lab.

    Techniques: Labeling, Stable Transfection, Expressing, Live Cell Imaging, Transfection, Imaging

    ( A ) Volcano plot of the TMED9 interactome (Myc-TMED9 + MUC1-fs) shows preferential binding of TMED9 (green dot) to COPI proteins (red dots), but not to COPII proteins (purple dots), which did not pass the significance threshold for enrichment compared to the negative control (empty vector + MUC1-fs). ( B ) Myc-tagged TMED9 co-IP’ed. COPB2 (COPI protein) but not SEC13 (COPII protein). β-Actin is shown as a loading control in the lysate and as a negative control in the pulldown. Blots are representative of three independent experiments. ( C ) Myc-tagged TMED9 mutant R223E coimmunoprecipitated less endogenous COPB2 than WT TMED9. ( D ) GFP-tagged SEC23a coimmunoprecipitated mutant and WT TMED9. ** P < 0.05. ns., not significant.

    Journal: Science Advances

    Article Title: Molecular basis of TMED9 oligomerization and entrapment of misfolded protein cargo in the early secretory pathway

    doi: 10.1126/sciadv.adp2221

    Figure Lengend Snippet: ( A ) Volcano plot of the TMED9 interactome (Myc-TMED9 + MUC1-fs) shows preferential binding of TMED9 (green dot) to COPI proteins (red dots), but not to COPII proteins (purple dots), which did not pass the significance threshold for enrichment compared to the negative control (empty vector + MUC1-fs). ( B ) Myc-tagged TMED9 co-IP’ed. COPB2 (COPI protein) but not SEC13 (COPII protein). β-Actin is shown as a loading control in the lysate and as a negative control in the pulldown. Blots are representative of three independent experiments. ( C ) Myc-tagged TMED9 mutant R223E coimmunoprecipitated less endogenous COPB2 than WT TMED9. ( D ) GFP-tagged SEC23a coimmunoprecipitated mutant and WT TMED9. ** P < 0.05. ns., not significant.

    Article Snippet: The SEC23A plasmid was ordered from Addgene, pEGFP-Sec23A (Addgene, catalog: 66609).

    Techniques: Binding Assay, Negative Control, Plasmid Preparation, Control, Mutagenesis