Anti-phospho-Neuroligin-1/NL-1 (Thr739)

88.93/87.09 kDa (human isoform 1/2), 89.12 kDa (mouse isoform 1), and 89.26/85.96/88.23/87.00 kDa (rat isoform 1/2/3/4) calculated (Signal peptide a.a. 1-45 removed). ~120 kDa reported (Bemben, M.A., et al. (2014). Nat. Neurosci. 17(1):56-64). Uncharacterized band(s) may appear in some lysates.

Neuroligin-1 (UniProt Q99K10; also known as NL-1) is encoded by the Nlgn1 (also known as Kiaa1070) gene (Gene ID 192167) in murine species. Neuroligins are single-transmembrane postsynaptic proteins that contain a large extracellular acetylcholinesterase-like domain and a cytoplasmic tail (c-tail) with many protein-protein interaction domains. Neuroligin-1 (NL-1) is localized predominantly to excitatory synapses and NL-2 is confined primarily to inhibitory synapses, while NL-3 is found among both types of synapses and NL-4 is enriched at glycinergic synapses. NL-1 is involved in diverse forms of synaptic plasticity across species. NL-1 overexpression enhances evoked excitatory postsynaptic currents (EPSCs) and synaptogenesis, while chronic blockade of NMDA receptors (NMDARs) or CaMKII diminishes NL-1–mediated synaptic potentiation. CaMKII is shown to phosphorylate NL-1 on Thr739 and NL-1 T739A mutation markedly reduces the basal and activity-induced NL-1 surface expression. NL-1 is produced with a signal peptide (a.a. 1-45) that is posttranslationally removed to yield the mature protein with a large extracellular domain (a.a. 46-697), followed by a transmembrane segment (a.a. 698-718) and a cytoplasmic tail (a.a. 719-843).

Epitope: pThr739.

Linear peptide corresponding to a mouse Neuroligin-1 sequence containing phosphorylated Thr739.

Anti-Neuroligin Antibody is an antibody against phospho-Neuroligin-1/NL-1 (Thr739) for use in Western Blotting, Immunoprecipitation.

Research Category
Neuroscience

Research Sub Category
Synapse & Synaptic Biology

Western Blotting Analysis: A representative lot detected CaMKII-catalyzed Thr739 phosphorylation of mouse NL-1 C-terminal fragment GST fusion in a kinase assay (Bemben, M.A., et al. (2014). Nat. Neurosci. 17(1):56-64).
Western Blotting Analysis: A representative lot specifically detected the target phospho-band in lysates from transfected cells co-expressing constitutively active CaMKII (T286D) and wild-type, but not T739A mutan, NL-1 (Bemben, M.A., et al. (2014). Nat. Neurosci. 17(1):56-64).
Western Blotting Analysis: A representative lot detected an enhanced NL-1 Thr730 phosphorylation in cultured embryonic rat cortical neurons upon GABA(A) receptor antagonist BCC (Cat. No. 505875) treatment. Co-treatment with NMDAR and AMPAR antagonists blocked the effect seen with BCC treatment alone (Bemben, M.A., et al. (2014). Nat. Neurosci. 17(1):56-64).
Immunoprecipitation Analysis: A representative lot immunoprecipitated a ~120 kDa Thr739-phosphorylated NL-1 from wild-type, but not Nlgn1-knockout, mouse brain tissue lysates (Bemben, M.A., et al. (2014). Nat. Neurosci. 17(1):56-64).
Immunoprecipitation Analysis: A representative lot immunoprecipitated less Thr739-phosphorylated NL-1 from visual cortex of postnatal mice following a 5-day dark-rearing (DR) period (no light from P21-P26). A two-hour light exposure after the 5-day DR period restored visual cortex NL-1 phosphorylation the control level (Bemben, M.A., et al. (2014). Nat. Neurosci. 17(1):56-64).

This polyclonal antibody specifically detected the target phospho-band in lysates from transfected cells co-expressing constitutively active CaMKII (T286D) and wild-type, but not T739A mutan, NL-1. The Thr739 designation is based on murine & rat NL-1 sequence (UniProt Q99K10 & Q62765). The equivalent site in human NL-1 is Thr736 (UniProt Q8N2Q7). The target phosphorylation site is present in both human spliced isoforms (Thr719 of isoform 2) and all four rat spliced isoforms (Thr710/730/719 of isoform 2/3/4), but is missing in mouse spliced isoform 2.

Affinity purified.

Purified rabbit polyclonal antibody in buffer containing PBS with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Evaluated by Western Blotting of CaMKII-catalyzed phosphorylation of mouse NL-1 GST fusion.

Western Blotting Analysis: 0.2 µg/mL of this antibody detected CaMKII-catalyzed Thr739 phosphorylation of 6 µg mouse NL-1 C-terminal fragment GST fusion in a kinase assay.

Concentration: Please refer to lot specific datasheet.

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