ABN1707

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

from rabbit, purified by affinity chromatography

• Synonym(s): Neuroligin-1, Thr739-phosphorylated, NL-1, Thr739-phosphorylated
• UNSPSC Code: 12352203
• eCl@ss: 32160702

Properties

• biological source: rabbit
• antibody form: affinity isolated antibody
• antibody product type: primary antibodies
• clone: polyclonal
• purified by: affinity chromatography
• species reactivity: mouse, rat
• species reactivity (predicted by homology): bovine (based on 100% sequence homology), rhesus macaque (based on 100% sequence homology), human (based on 100% sequence homology), chicken (based on 100% sequence homology)
• technique(s): immunoprecipitation (IP): suitable; western blot: suitable
• NCBI accession no.: NP_619607
• UniProt accession no.: Q99K10
• target post-translational modification: phosphorylation (pThr739 )
• Gene Information: mouse ... Nlgn1(192167)

Description

General description

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).

Specificity

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.

Immunogen

Epitope: pThr739.

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

Application

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).

Quality

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.

Target description

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.

Physical form

Affinity purified.

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

Storage and Stability

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

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class: 12 - Non Combustible Liquids
• wgk_germany: WGK 2
• flash_point_f: Not applicable
• flash_point_c: Not applicable