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  • 94
    Alomone Labs agc004
    Agc004, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Alomone Labs guinea pig anti glua1
    a ) Diagram of SYNPLA targeting presynaptic myc-NRXN and. cLTP induces mobilization of <t>GluA1</t> from an extrasynaptic pool and insertion into the post-synaptic density, decreasing the distance between the targeted proteins and permitting PLA. b ) Representative images of SYNPLA reactions performed on myc-NRXN expressing cultured rat hippocampal neurons at 14 days in vitro for the indicated conditions; PLA shown in gray. Scale bar: 10 μm. c ) Quantification of a single SYNPLA experiment (as in f ). The number of SYNPLA puncta detected in each field of view (round) and the average (square) ± SEM across all fields of view are shown; *** p<0.001, 1-way ANOVA followed by Tukey-Kramer post-hoc test. d ) Quantification of six independent experiments (indicated by different colors); the average number of SYNPLA puncta across 10-12 fields of view for each experiment (circles, normalized to CTRL) and the average PLA signal across experiments (square) ± SEM are shown; ** p<0.01, 1-way ANOVA (with Tukey-Kramer post-hoc test). e ) Diagram of SYNPLA in rat organotypic hippocampal slices. Sindbis virus expressing myc-NRXN is injected in the presynaptic CA3 region SYNPLA between myc-NRXN and endogenous GluA1 is measured in the postsynaptic CA1 region. f ) Representative images of SYNPLA under the indicated conditions in region CA1 of organotypic slice cultures. Scale bar: 10 μm. g ) Quantification of SYNPLA puncta in organotypic slices, from three independent experiments (indicated with different colors, 4 slices per condition (one circle per slice)); squares indicate average ± SEM across experiments; *** p<0.001, paired t-test.
    Guinea Pig Anti Glua1, supplied by Alomone Labs, 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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    94
    Alomone Labs rabbit polyclonal anti ampa receptor 1
    (A and B) Upper: representative images of individual endogenous <t>GluA1</t> (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.
    Rabbit Polyclonal Anti Ampa Receptor 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti ampa receptor 1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    a ) Diagram of SYNPLA targeting presynaptic myc-NRXN and. cLTP induces mobilization of GluA1 from an extrasynaptic pool and insertion into the post-synaptic density, decreasing the distance between the targeted proteins and permitting PLA. b ) Representative images of SYNPLA reactions performed on myc-NRXN expressing cultured rat hippocampal neurons at 14 days in vitro for the indicated conditions; PLA shown in gray. Scale bar: 10 μm. c ) Quantification of a single SYNPLA experiment (as in f ). The number of SYNPLA puncta detected in each field of view (round) and the average (square) ± SEM across all fields of view are shown; *** p<0.001, 1-way ANOVA followed by Tukey-Kramer post-hoc test. d ) Quantification of six independent experiments (indicated by different colors); the average number of SYNPLA puncta across 10-12 fields of view for each experiment (circles, normalized to CTRL) and the average PLA signal across experiments (square) ± SEM are shown; ** p<0.01, 1-way ANOVA (with Tukey-Kramer post-hoc test). e ) Diagram of SYNPLA in rat organotypic hippocampal slices. Sindbis virus expressing myc-NRXN is injected in the presynaptic CA3 region SYNPLA between myc-NRXN and endogenous GluA1 is measured in the postsynaptic CA1 region. f ) Representative images of SYNPLA under the indicated conditions in region CA1 of organotypic slice cultures. Scale bar: 10 μm. g ) Quantification of SYNPLA puncta in organotypic slices, from three independent experiments (indicated with different colors, 4 slices per condition (one circle per slice)); squares indicate average ± SEM across experiments; *** p<0.001, paired t-test.

    Journal: bioRxiv

    Article Title: SYNPLA: A synapse-specific method for identifying learning-induced synaptic plasticity loci

    doi: 10.1101/473314

    Figure Lengend Snippet: a ) Diagram of SYNPLA targeting presynaptic myc-NRXN and. cLTP induces mobilization of GluA1 from an extrasynaptic pool and insertion into the post-synaptic density, decreasing the distance between the targeted proteins and permitting PLA. b ) Representative images of SYNPLA reactions performed on myc-NRXN expressing cultured rat hippocampal neurons at 14 days in vitro for the indicated conditions; PLA shown in gray. Scale bar: 10 μm. c ) Quantification of a single SYNPLA experiment (as in f ). The number of SYNPLA puncta detected in each field of view (round) and the average (square) ± SEM across all fields of view are shown; *** p<0.001, 1-way ANOVA followed by Tukey-Kramer post-hoc test. d ) Quantification of six independent experiments (indicated by different colors); the average number of SYNPLA puncta across 10-12 fields of view for each experiment (circles, normalized to CTRL) and the average PLA signal across experiments (square) ± SEM are shown; ** p<0.01, 1-way ANOVA (with Tukey-Kramer post-hoc test). e ) Diagram of SYNPLA in rat organotypic hippocampal slices. Sindbis virus expressing myc-NRXN is injected in the presynaptic CA3 region SYNPLA between myc-NRXN and endogenous GluA1 is measured in the postsynaptic CA1 region. f ) Representative images of SYNPLA under the indicated conditions in region CA1 of organotypic slice cultures. Scale bar: 10 μm. g ) Quantification of SYNPLA puncta in organotypic slices, from three independent experiments (indicated with different colors, 4 slices per condition (one circle per slice)); squares indicate average ± SEM across experiments; *** p<0.001, paired t-test.

    Article Snippet: For Supplementary Fig. 2 different primary antibodies were used as negative controls: guinea pig anti-GluA1 (AGP-009, Alomone labs), rabbit anti-cFos (Cell signaling, #2250); all 1:100 dilution.

    Techniques: Expressing, Cell Culture, In Vitro, Injection

    a ) Diagram of a general PLA reaction. Note that A and B must be sufficiently close to permit complementary binding and ligation of ABo1 and ABo2 and subsequent amplification to make a rolony. b ) Diagram of PLA targeting recombinant presynaptic myc-NRXN and postsynaptic HA-NLGN. c ) PLA rolonies (white dots, right) formed between postsynaptic cultured mouse hippocampal neuron expressing HA-NLGN + mCherry (red; middle) and co-cultured presynaptic neurons expressing myc-NRXN + GFP (green; left). Scale bar: 10 μm. d ) PLA puncta in each sample (circles) and average (squares) ± SEM for indicated expression and days in vitro (color indicates data acquired on the same day). *** p<0.001, 2-way ANOVA (see Methods); * p<0.05, 1-way ANOVA (with Tukey-Kramer post-hoc test). e ) PLA reaction between endogenous GluA1 and recombinant myc in cultured rat hippocampal neurons (exposed to cLTP, see ) expressing myc-NRXN and cytosolic fluorescent protein (green), subsequently immunostained for endogenous PSD-95 (red). Scale bars: 10 μm (left); 5 μm (right). f ) PLA puncta localize to synapses. In PSD-95 and myc-NRXN channels (from e , right), pixels were set to zero (masked) for values below a progressively increasing threshold (x axis). Puncta (PLA or randomly placed) display colocalization if non-zero pixels exist within 0.14 μm in both thresholded channels. For indicated threshold, ∼95% of PLA, while only ∼50% of randomly placed (mean ± SEM of 30 placements) puncta colocalized with pre- and post-synaptic markers. See Supplementary Fig. 3 and Methods for details.

    Journal: bioRxiv

    Article Title: SYNPLA: A synapse-specific method for identifying learning-induced synaptic plasticity loci

    doi: 10.1101/473314

    Figure Lengend Snippet: a ) Diagram of a general PLA reaction. Note that A and B must be sufficiently close to permit complementary binding and ligation of ABo1 and ABo2 and subsequent amplification to make a rolony. b ) Diagram of PLA targeting recombinant presynaptic myc-NRXN and postsynaptic HA-NLGN. c ) PLA rolonies (white dots, right) formed between postsynaptic cultured mouse hippocampal neuron expressing HA-NLGN + mCherry (red; middle) and co-cultured presynaptic neurons expressing myc-NRXN + GFP (green; left). Scale bar: 10 μm. d ) PLA puncta in each sample (circles) and average (squares) ± SEM for indicated expression and days in vitro (color indicates data acquired on the same day). *** p<0.001, 2-way ANOVA (see Methods); * p<0.05, 1-way ANOVA (with Tukey-Kramer post-hoc test). e ) PLA reaction between endogenous GluA1 and recombinant myc in cultured rat hippocampal neurons (exposed to cLTP, see ) expressing myc-NRXN and cytosolic fluorescent protein (green), subsequently immunostained for endogenous PSD-95 (red). Scale bars: 10 μm (left); 5 μm (right). f ) PLA puncta localize to synapses. In PSD-95 and myc-NRXN channels (from e , right), pixels were set to zero (masked) for values below a progressively increasing threshold (x axis). Puncta (PLA or randomly placed) display colocalization if non-zero pixels exist within 0.14 μm in both thresholded channels. For indicated threshold, ∼95% of PLA, while only ∼50% of randomly placed (mean ± SEM of 30 placements) puncta colocalized with pre- and post-synaptic markers. See Supplementary Fig. 3 and Methods for details.

    Article Snippet: For Supplementary Fig. 2 different primary antibodies were used as negative controls: guinea pig anti-GluA1 (AGP-009, Alomone labs), rabbit anti-cFos (Cell signaling, #2250); all 1:100 dilution.

    Techniques: Binding Assay, Ligation, Amplification, Recombinant, Cell Culture, Expressing, In Vitro

    (A and B) Upper: representative images of individual endogenous GluA1 (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.

    Journal: Cell reports

    Article Title: CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X

    doi: 10.1016/j.celrep.2022.110853

    Figure Lengend Snippet: (A and B) Upper: representative images of individual endogenous GluA1 (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.

    Article Snippet: Rabbit polyclonal anti-AMPA Receptor 1 (GluR1)/GluA1 , Alomone Labs , Cat# AGC-004; RRID:AB_2039878.

    Techniques: Immunofluorescence, Marker

    (A–D) Representative images of endogenous single mRNAs molecules encoding GluA1 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (A) quantified in (B). (C) Higher-magnification images of boxed region in (A) quantified in (D). (E–H) Representative images of endogenous single mRNAs molecules encoding GluA2 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (E) quantified in (F). (G) Higher-magnification images of boxed region in (E) quantified in (H) (GluA1, WT: 101 dendrites, 35 neurons; KO: 134 dendrites, 63 neurons, n = 4 per group; GluA2, WT: 190 dendrites, 82 neurons; KO: 99 dendrites, 82 neurons, n = 4 per group). (I) Representative images of a dual FISH experiment with probes targeting either the GluA2 coding (upper left) or intronic (upper right) sequences. (J) Representative images showing the number of GluA1 and GluA2 mRNA molecules at transcription hotspots in the nucleus of neurons from WT and Fmr1 KO mice. (K) Summary data of images like those illustrated in (J) (GluA1, WT: 35 neurons; KO: 63 neurons; GluA2, WT: 82 neurons, KO: 86 neurons, n = 4). (L) Summary qRT-PCR data from whole hippocampus showing GluA1 (upper) and GluA2 (lower) mRNA expression (n = 4 per group). For all the quantitative graphs: Data are mean ± SEM. *p < 0.05; **p < 0.01.

    Journal: Cell reports

    Article Title: CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X

    doi: 10.1016/j.celrep.2022.110853

    Figure Lengend Snippet: (A–D) Representative images of endogenous single mRNAs molecules encoding GluA1 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (A) quantified in (B). (C) Higher-magnification images of boxed region in (A) quantified in (D). (E–H) Representative images of endogenous single mRNAs molecules encoding GluA2 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (E) quantified in (F). (G) Higher-magnification images of boxed region in (E) quantified in (H) (GluA1, WT: 101 dendrites, 35 neurons; KO: 134 dendrites, 63 neurons, n = 4 per group; GluA2, WT: 190 dendrites, 82 neurons; KO: 99 dendrites, 82 neurons, n = 4 per group). (I) Representative images of a dual FISH experiment with probes targeting either the GluA2 coding (upper left) or intronic (upper right) sequences. (J) Representative images showing the number of GluA1 and GluA2 mRNA molecules at transcription hotspots in the nucleus of neurons from WT and Fmr1 KO mice. (K) Summary data of images like those illustrated in (J) (GluA1, WT: 35 neurons; KO: 63 neurons; GluA2, WT: 82 neurons, KO: 86 neurons, n = 4). (L) Summary qRT-PCR data from whole hippocampus showing GluA1 (upper) and GluA2 (lower) mRNA expression (n = 4 per group). For all the quantitative graphs: Data are mean ± SEM. *p < 0.05; **p < 0.01.

    Article Snippet: Rabbit polyclonal anti-AMPA Receptor 1 (GluR1)/GluA1 , Alomone Labs , Cat# AGC-004; RRID:AB_2039878.

    Techniques: Quantitative RT-PCR, Expressing

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X

    doi: 10.1016/j.celrep.2022.110853

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Rabbit polyclonal anti-AMPA Receptor 1 (GluR1)/GluA1 , Alomone Labs , Cat# AGC-004; RRID:AB_2039878.

    Techniques: shRNA, Recombinant, Sequencing, Binding Assay, Plasmid Preparation, Software