rabbit anti glud1501600 af647  (Bioss)

Journal: Nature Communications

Article Title: Neurexin-3 subsynaptic densities are spatially distinct from Neurexin-1 and essential for excitatory synapse nanoscale organization in the hippocampus

doi: 10.1038/s41467-023-40419-2

Figure Lengend Snippet: a Representative 3D dSTORM reconstruction of LRRTM2 (cyan) and Homer1 (red). b LRRTM2 Homer1 synaptic abundance n = 232 synapses from 3 independent experiments (left) and number of SSDs n = 151 synapses from 3 independent experiments (right). LRRTM2 localization distance ( c ) and SSD distance ( d ) to Homer1 centroid. The mean and 95% confidence interval of Poisson fitted distribution are indicated on the graph. e Representative 3D dSTORM reconstruction of LRRTM2 (cyan) and Nrxn1 (left) or Nrxn3 (right; red) at Homer1+ synapse (green) in an enface view. f Nrxn1 is farther from LRRTM2 SSDs than Nrxn3, p = 0.0135; Mann–Whitney test (two-tailed). g Representative 3D dSTORM reconstruction of GluD1 (cyan) and Homer1 (red) in an enface view. h GluD1 Homer1 synaptic abundance n = 196 synapses from 3 independent experiments (left) and number of SSDs n = 134 synapses from 3 independent experiments (right). i – j GluD1 localization distance ( g ) and GluD1 SSD distance ( h ) from the Homer1 centroid. The mean and 95% confidence interval of the Poisson fitted distribution indicated on the graph. k Representative 3D dSTORM reconstruction of GluD1 (cyan) and Nrxn1 (left) or Nrxn3 (right; red) at Homer1+ synapse (green) in an enface view. l Nrxn1 is closer to GluD1 SSDs than Nrxn1, p = 0.0314; Mann–Whitney test (two-tailed). Number of synapses and independent experiments are indicated in the figure. Bar graphs and line graphs: average ± SEM. Violin plots: median ± upper and lower quartiles. * p < 0.05. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies that detected intracellular protein sequences were then diluted in NGS blocking buffer incubated with the sample overnight at 4 C. Primary antibody dilutions were as follows: mouse anti-PSD-95 (ThermoFisher Cat# MA1-045; conjugated with kit; validated by relative expression by ThermoFisher) 1:50, rabbit anti-RIM (Sysy Cat# 140 003; conjugated with kit; validated by knockout ) 1:25, rabbit anti-GluD1 501-600 AF647 (Bioss Cat# bs-12095R-A647; validated by flow cytometry), guinea pig anti-GluD1 895-932 (Frontier Institute Cat# GluD1C-GP-Af840; indirectly stained; validated by knockout ) 1:1000, and chicken anti-Homer1 (Sysy Cat# 160 006; indirectly stained; specific for Homer1 ) 1:1000.

Techniques: MANN-WHITNEY, Two Tailed Test

Journal: Nature Communications

Article Title: Neurexin-3 subsynaptic densities are spatially distinct from Neurexin-1 and essential for excitatory synapse nanoscale organization in the hippocampus

doi: 10.1038/s41467-023-40419-2

Figure Lengend Snippet: We find that Nrxn3 controls excitatory synapse nano-organization in hippocampus and that Nrxn1 and Nrxn3 localize in discrete, non-overlapping SSDs that are preferentially opposite GluD1 and LRRTM2 SSDs respectively. Nrxn3 deletion results in decreased AMPAR SSDs per synapse as well as a decrease in the relative number of AMPARs in SSDs.

Article Snippet: Primary antibodies that detected intracellular protein sequences were then diluted in NGS blocking buffer incubated with the sample overnight at 4 C. Primary antibody dilutions were as follows: mouse anti-PSD-95 (ThermoFisher Cat# MA1-045; conjugated with kit; validated by relative expression by ThermoFisher) 1:50, rabbit anti-RIM (Sysy Cat# 140 003; conjugated with kit; validated by knockout ) 1:25, rabbit anti-GluD1 501-600 AF647 (Bioss Cat# bs-12095R-A647; validated by flow cytometry), guinea pig anti-GluD1 895-932 (Frontier Institute Cat# GluD1C-GP-Af840; indirectly stained; validated by knockout ) 1:1000, and chicken anti-Homer1 (Sysy Cat# 160 006; indirectly stained; specific for Homer1 ) 1:1000.

Techniques: