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psd95 trfp  (Addgene inc)


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    Addgene inc psd95 trfp

    Psd95 Trfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/psd95 trfp/product/Addgene inc
    Average 93 stars, based on 16 article reviews
    psd95 trfp - by Bioz Stars, 2026-04
    93/100 stars

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    1) Product Images from "Endogenous Syngap1 alpha splice forms promote cognitive function and seizure protection"

    Article Title: Endogenous Syngap1 alpha splice forms promote cognitive function and seizure protection

    Journal: eLife

    doi: 10.7554/eLife.75707


    Figure Legend Snippet:

    Techniques Used: In Vitro, Plasmid Preparation, Sequencing, Virus, Bicinchoninic Acid Protein Assay, Software



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    Figure 5. Validation of SynGAP PDZ binding motif (PBM) mutations and construction of the Syngap1PBM mouse line. (A) Schematic diagram for exon map and alternative use of Exon21 in Syngap1 gene. Exon21b encodes for α1 isoform. Exon 21 a encodes for α2 isoform. Point mutations indicated in red alter exon 21b coding sequence without influencing exon21a open reading frame. (B) Schematics of SynGAPα1 and <t>PSD95</t> domain structure and the location of point mutations. (C) Illustrations of constructs expressed in HeLa cells to study PDZ-dependent interaction between SynGAP and PSD95.
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    Journal: eLife

    Article Title: Endogenous Syngap1 alpha splice forms promote cognitive function and seizure protection

    doi: 10.7554/eLife.75707

    Figure Lengend Snippet:

    Article Snippet: HeLa cells were plated on glass coverslips and transfected with PSD95-tRFP (Plasmid #52671, Addgene) and/or EGFP-tagged SynGAP C-terminal constructs EGFP-CCα1 or EGFP-CCPBM plasmids (made in house) were co-transfected into HeLa cells using lipofectamine 2000 according to manufacturer instructions.

    Techniques: In Vitro, Plasmid Preparation, Sequencing, Virus, Bicinchoninic Acid Protein Assay, Software

    Figure 5. Validation of SynGAP PDZ binding motif (PBM) mutations and construction of the Syngap1PBM mouse line. (A) Schematic diagram for exon map and alternative use of Exon21 in Syngap1 gene. Exon21b encodes for α1 isoform. Exon 21 a encodes for α2 isoform. Point mutations indicated in red alter exon 21b coding sequence without influencing exon21a open reading frame. (B) Schematics of SynGAPα1 and PSD95 domain structure and the location of point mutations. (C) Illustrations of constructs expressed in HeLa cells to study PDZ-dependent interaction between SynGAP and PSD95.

    Journal: eLife

    Article Title: Endogenous Syngap1 alpha splice forms promote cognitive function and seizure protection

    doi: 10.7554/elife.75707

    Figure Lengend Snippet: Figure 5. Validation of SynGAP PDZ binding motif (PBM) mutations and construction of the Syngap1PBM mouse line. (A) Schematic diagram for exon map and alternative use of Exon21 in Syngap1 gene. Exon21b encodes for α1 isoform. Exon 21 a encodes for α2 isoform. Point mutations indicated in red alter exon 21b coding sequence without influencing exon21a open reading frame. (B) Schematics of SynGAPα1 and PSD95 domain structure and the location of point mutations. (C) Illustrations of constructs expressed in HeLa cells to study PDZ-dependent interaction between SynGAP and PSD95.

    Article Snippet: Reagent type (species) or resource Designation Source or reference Identifiers Additional information Biological sample (Mus musculus) Mouse primary forebrain neurons This study 18–21 days in vitro Biological sample (Mus musculus) Cortical and hippocampal tissue Multiple strains (this study) Male and female Strain, strain background (Mus musculus) IRES- TD Spicer et al., 2018 C57BL/6 J background Strain, strain background (Mus musculus) Beta KO (β*) This study C57BL/6 J background Strain, strain background (Mus musculus) PBM This study C57BL/6 J background Cell line (Homo sapiens) HeLa Farzan Lab (Scripps Research) Cell line (Homo sapiens) H293T Kissil Lab (Scripps Research) Genetic reagent (Rattus norvegicus) PSD95- tRFP Addgene #52,671 Plasmid Genetic reagent (Mus musculus) EGFP- CCα1 This study Plasmid Genetic reagent (Mus musculus) EGFP- CCPBM This study Plasmid Genetic reagent (Mus musculus) EGFP- SynGAPα1 This study Plasmid Genetic reagent (Mus musculus) EGFP- SynGAPα1PBM This study Plasmid Antibody Rabbit polyclonal antibody Thermo PA1- 046 Anti- total SynGAP (1:1000) Antibody Rabbit polyclonal antibody Millipore 06–900 Anti- SynGAPα1 (1:1000) Antibody Rabbit monoclonal antibody Abcam ab77235 Anti- SynGAPα2 (1:1000) Antibody Rabbit polyclonal antibody Araki et al., 2020 Huganir Lab (JHU) Anti- SynGAPβ (1:1000) Antibody Mouse monoclonal antibody Thermo MA1- 045 Anti- PSD95 (1:2000) Antibody Rabbit polyclonal antibody Novus NB300- 653 Anti- Synaptophysin (1:1000) Antibody Mouse monoclonal antibody CST 9,106 Anti- phosphoERK (1:1000) Antibody Mouse monoclonal antibody CST 4,696 Anti- ERK (1:1000) Antibody Mouse monoclonal antibody Millipore MAB2263 Anti- GluA1 N- term. (1:500) Antibody Rabbit polyclonal antibody Millipore Ab9876 Anti- TARP (1:500) Antibody Rabbit polyclonal antibody Thermo Pierce PA521097 Anti- LRRTM2 (1:1000) Sequence- based reagent IRES- TD Genotyping Primer Fw IDT AGAT CCAC CAGG CCCT GAA Sequence- based reagent IRES- TD Genotyping Primer Rev IDT GTCT TGAA CTCC ACCA GGTA GTG Sequence- based reagent PBM Genotyping Primer Fw IDT CTGG TTCA AAGG CTCC TGGT A Sequence- based reagent PBM Genotyping Primer Rev IDT CTGT TTGT TTCT CACC TCCA GGAA Kilinc et al. eLife 2022;11:e75707.

    Techniques: Biomarker Discovery, Binding Assay, Sequencing, Construct

    Figure 7. Characterization of native PSD95 complexes from Syngap1PBM animals. (A) Volcano plot demonstrating the label-free quantitative mass- spectrometry profile of the logarithmic difference in protein levels in the immunoprecipitated PSD95 complexes derived from DIV21 +/+ and PBM/ PBM cultures in inactivated state. Only Gpm6a (shown in black) was significantly altered beyond p > 0.001 cutoff. Blue dots represent proteins with type 1 PDZ-ligands. Green dots represent DLG family proteins. p Values were calculated via t-test for each protein. Samples were derived from individual cultures (4 per genotype) which are immunoprecipitated separately. Log2FC was calculated as ratio of PBM/PBM over +/+. (B) List of proteins that are differentially expressed beyond p > 0.05 cutoff. Note that Iqseq2 and Dlgap3 are PDZ-binding proteins. (C) Mass-spectrometry profile of type-1 PDZ binding motif containing proteins in immunoprecipitated PSD95 complex in +/+ vs PBM/PBM inactivated cultures. (D) Western blots showing relative expression of TARPs and Lrrtm2 in PSD fractions from adult hippocampi in +/+ vs PBM/PBM. (E–G) Quantifications of (D). (E) TARPg8 t(6)=0.6961, P = 0.5124. (F) TARPg2-4 t(6)=0.7924, p = 0.4583 (G) Lrrtm2 t(6)=0.5542, p = 0.5995. Each sample represents hippocampi pooled from 2 mice. (H) Comparison of PSD95 and SynGAP IP complexes as reported by Li et al., 2017 and Wilkinson et al., 2017. Note that PSD95 and SynGAP complexes share diverse range of components involving cytoskeletal and scaffolding proteins.

    Journal: eLife

    Article Title: Endogenous Syngap1 alpha splice forms promote cognitive function and seizure protection

    doi: 10.7554/elife.75707

    Figure Lengend Snippet: Figure 7. Characterization of native PSD95 complexes from Syngap1PBM animals. (A) Volcano plot demonstrating the label-free quantitative mass- spectrometry profile of the logarithmic difference in protein levels in the immunoprecipitated PSD95 complexes derived from DIV21 +/+ and PBM/ PBM cultures in inactivated state. Only Gpm6a (shown in black) was significantly altered beyond p > 0.001 cutoff. Blue dots represent proteins with type 1 PDZ-ligands. Green dots represent DLG family proteins. p Values were calculated via t-test for each protein. Samples were derived from individual cultures (4 per genotype) which are immunoprecipitated separately. Log2FC was calculated as ratio of PBM/PBM over +/+. (B) List of proteins that are differentially expressed beyond p > 0.05 cutoff. Note that Iqseq2 and Dlgap3 are PDZ-binding proteins. (C) Mass-spectrometry profile of type-1 PDZ binding motif containing proteins in immunoprecipitated PSD95 complex in +/+ vs PBM/PBM inactivated cultures. (D) Western blots showing relative expression of TARPs and Lrrtm2 in PSD fractions from adult hippocampi in +/+ vs PBM/PBM. (E–G) Quantifications of (D). (E) TARPg8 t(6)=0.6961, P = 0.5124. (F) TARPg2-4 t(6)=0.7924, p = 0.4583 (G) Lrrtm2 t(6)=0.5542, p = 0.5995. Each sample represents hippocampi pooled from 2 mice. (H) Comparison of PSD95 and SynGAP IP complexes as reported by Li et al., 2017 and Wilkinson et al., 2017. Note that PSD95 and SynGAP complexes share diverse range of components involving cytoskeletal and scaffolding proteins.

    Article Snippet: Reagent type (species) or resource Designation Source or reference Identifiers Additional information Biological sample (Mus musculus) Mouse primary forebrain neurons This study 18–21 days in vitro Biological sample (Mus musculus) Cortical and hippocampal tissue Multiple strains (this study) Male and female Strain, strain background (Mus musculus) IRES- TD Spicer et al., 2018 C57BL/6 J background Strain, strain background (Mus musculus) Beta KO (β*) This study C57BL/6 J background Strain, strain background (Mus musculus) PBM This study C57BL/6 J background Cell line (Homo sapiens) HeLa Farzan Lab (Scripps Research) Cell line (Homo sapiens) H293T Kissil Lab (Scripps Research) Genetic reagent (Rattus norvegicus) PSD95- tRFP Addgene #52,671 Plasmid Genetic reagent (Mus musculus) EGFP- CCα1 This study Plasmid Genetic reagent (Mus musculus) EGFP- CCPBM This study Plasmid Genetic reagent (Mus musculus) EGFP- SynGAPα1 This study Plasmid Genetic reagent (Mus musculus) EGFP- SynGAPα1PBM This study Plasmid Antibody Rabbit polyclonal antibody Thermo PA1- 046 Anti- total SynGAP (1:1000) Antibody Rabbit polyclonal antibody Millipore 06–900 Anti- SynGAPα1 (1:1000) Antibody Rabbit monoclonal antibody Abcam ab77235 Anti- SynGAPα2 (1:1000) Antibody Rabbit polyclonal antibody Araki et al., 2020 Huganir Lab (JHU) Anti- SynGAPβ (1:1000) Antibody Mouse monoclonal antibody Thermo MA1- 045 Anti- PSD95 (1:2000) Antibody Rabbit polyclonal antibody Novus NB300- 653 Anti- Synaptophysin (1:1000) Antibody Mouse monoclonal antibody CST 9,106 Anti- phosphoERK (1:1000) Antibody Mouse monoclonal antibody CST 4,696 Anti- ERK (1:1000) Antibody Mouse monoclonal antibody Millipore MAB2263 Anti- GluA1 N- term. (1:500) Antibody Rabbit polyclonal antibody Millipore Ab9876 Anti- TARP (1:500) Antibody Rabbit polyclonal antibody Thermo Pierce PA521097 Anti- LRRTM2 (1:1000) Sequence- based reagent IRES- TD Genotyping Primer Fw IDT AGAT CCAC CAGG CCCT GAA Sequence- based reagent IRES- TD Genotyping Primer Rev IDT GTCT TGAA CTCC ACCA GGTA GTG Sequence- based reagent PBM Genotyping Primer Fw IDT CTGG TTCA AAGG CTCC TGGT A Sequence- based reagent PBM Genotyping Primer Rev IDT CTGT TTGT TTCT CACC TCCA GGAA Kilinc et al. eLife 2022;11:e75707.

    Techniques: Mass Spectrometry, Immunoprecipitation, Derivative Assay, Binding Assay, Western Blot, Expressing, Comparison, Scaffolding