Recombinant Mouse NUCKS1 Protein

• Cat.No.: NUCKS1-10956M
• Product Overview: Recombinant Mouse NUCKS1 full length or partial length protein was expressed.

Specification

• Species: Mouse
• Source: Mammalian Cells
• Tag: His
• Form: Liquid or lyophilized powder
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method.
• Purity: >80%
• Notes: This item requires custom production and lead time is between 5-9 weeks. We can custom produce according to your specifications.
• Storage: Store it at +4 ºC for short term. For long term storage, store it at -20 ºC～-80 ºC.
• Storage Buffer: PBS buffer
• Publications:
  a-Endosulfine regulates amyloid b 42 via the modulation of neprilysin activity (2020)

Gene Information

• Gene Name: Nucks1 nuclear casein kinase and cyclin-dependent kinase substrate 1 [ Mus musculus ]
• Official Symbol: NUCKS1
• Gene ID: 98415
• mRNA Refseq: NM_001145804.1
• Protein Refseq: NP_001139276.1
• UniProt ID: Q80XU3

Citation

Somatostatin-evoked Aβ catabolism in the brain: Mechanistic involvement of α-endosulfine-K ATP channel pathway

Journal: Molecular Psychiatry    PubMed ID: 34737456    Data: 2021/11/4

Authors: Naoto Watamura, Naomasa Kakiya, Takaomi C. Saido

Article Snippet:NEP activity measurements were performed on primary neurons after 15–28 days of in vitro (DIV15–28) culture as previously described [ ].NEP activity measurements were performed on primary neurons after 15–28 days of in vitro (DIV15–28) culture as previously described [ ].. Somatostatin (Peptide institute 4023), TT232 (Tocris 3493), recombinant ENSA (abcam ab92932), recombinant NSG-1 (Creative BioMart NSG1–332H), recombinant NUCKS-1 (Creative BioMart NUCKS1–10956M) and diazoxide (Wako 364-98-7) were added as appropriate concentrations, and cells were incubated for a further 24 h. Neurons were then incubated with substrate mixture 50 μM suc-Ala-Ala-Phe-MCA (Sigma S8758), 10 nM benzyloxycarbonyl Z-Leu-Leu-Leucinal (Peptide institute 3175-V) and cOmplete EDTA-Free-Protease inhibitor (Roche Diagnostics 4693132) in 0.2 M MES buffer (pH6.5) with or without Thiorphan (Sigma T6031) for 1 h at 37 ?C.. Following this, 0.1 mM phosphoramidon (Peptide Institute 4082) and 0.1 mg/ml leucine aminopeptidase (Sigma L-5006) were added, and the reaction mixture was incubated at 37 ?C for a further 30 min. 7-Amino-4-methylcoumarin fluorescence was measured at excitation and emission wavelengths of 380 nm and 460 nm, respectively.Following this, 0.1 mM phosphoramidon (Peptide Institute 4082) and 0.1 mg/ml leucine aminopeptidase (Sigma L-5006) were added, and the reaction mixture was..

A-C. NEP activity after treatment of co-cultured cells with 1 μM somatostatin or TT232 for 24 h. A Cortical/hippocampal (Ctx&Hip) neurons ( n = 12 wells per treatment), ( B ) co-cultured neurons ( n = 10 wells per treatment), and ( C ) basal ganglia neurons ( n = 8 or 9 wells per treatment) were used. D – F NEP activity in co-cultured neurons after the replacement of the culture medium with conditioned media from ( E ) Ctx&Hip and ( F ) basal ganglia neurons treated with 1 μM somatostatin for 0–6 h. n = 6–10 wells per treatment in co-cultured neurons. G – K NEP activity of co-cultured neurons after replacement of the culture medium with separated conditioned media from Ctx&Hip neurons treated with SST or TT232. H , I 10 and ( J and K ) 30kDa centrifugal filters were used for the separation ( n = 7–10 for each group). NEP activity in co-cultured neurons after incubation with ( L ) ENSA, ( M ) NSG-1 and ( N ) NUCKS-1 recombinant proteins for 24 h. n = 8–10 wells per treatment in co-cultured neurons. Data represent the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 (one-way ANOVA with Dunnett’s post-hoc test).

α-Endosulfine regulates amyloid β 42 via the modulation of neprilysin activity

Journal: bioRxiv    Data: 2020/10/8

Authors: Watamura Naoto, Kakiya Naomasa, Saido Takaomi C.

Article Snippet:PrePrint: NEP activity measurements were performed on primary neurons after 15-28 days of in vitro (DIV15-28) culture as previously described .NEP activity measurements were performed on primary neurons after 15-28 days of in vitro (DIV15-28) culture as previously described .. Somatostatin (Peptide institute 4023), TT232 (Tocris 3493), recombinant ENSA (abcam ab92932), recombinant NSG-1 (Creative BioMart NSG1-332H), recombinant NUCKS-1 (Creative BioMart NUCKS1-10956M) and diazoxide (Wako 364-98-7) were added as appropriate concentrations, and cells were incubated for a further 24 hours.. Neurons were then incubated with substrate mixture (50 μM suc-Ala-Ala-Phe-MCA (Sigma S8758), 10 nM benzyloxycarbonyl Z-Leu-Leu-Leucinal (Peptide institute 3175-V) and cOmplete EDTA-Free-Protease inhibitor (Roche Diagnostics 4693132) in 0.2 M MES buffer (pH6.5) with or without Thiorphan (Sigma T6031) for 1 hour at 37°C.Neurons were then incubated with substrate mixture (50 μM suc-Ala-Ala-Phe-MCA (Sigma S8758), 10 nM benzyloxycarbonyl Z-Leu-Leu-Leucinal (Peptide institute 3175-V) and cOmplete EDTA-Free-Protease inhibitor (Roche Diagnostics 4693132) in 0.2 M MES buffer (pH6.5) with or without Thiorphan (Sigma T6031) for 1 hour at 37°C.

a-c, NEP activity after treatment of co-cultured cells with 1 μM somatostatin or TT232 for 24 hours. Cortical/hippocampal (Ctx&Hip) neurons (n = 12 wells per treatment), co-cultured neurons (n = 10 wells per treatment), and basal ganglia neurons (n = 8 or 9 wells per treatment) were used. d-f, NEP activity in co-cultured neurons after the replacement of the culture medium with conditioned media from Ctx&Hip and basal ganglia neurons treated with 1 μM somatostatin for 0-6 hours. n = 6-10 wells per treatment in co-cultured neurons. g-i, NEP activity in co-cultured neurons after incubation with ENSA, NSG-1 and NUCKS-1 recombinant proteins for 24 hours. n = 8-10 wells per treatment in co-cultured neurons. j-l, Immunoblotting of ENSA in 3-month-old WT, Sst 1 /Sst 4 dKO and Srif KO mice (n = 4 for each group). Values indicated in the graph show ENSA band intensities normalized to that of β-actin. m-p, Immunostaining of ENSA in the cortices and hippocampal CA1 and CA3 regions from 3-month-old WT, Sst 1 /Sst 4 dKO, and Srif KO mice (n = 6 for each group). Data represent the mean ±SEM. *P <0.05, **P <0.01, ***P <0.001 (one-way ANOVA with Dunnett’s post-hoc test).

a, Immunoblotting of NSG-1 or NUCKS-1 incubated with or without NEP and thiorphan for 24 hours at 37°C. The specific band is detected by Myc-tag antibody. b, Immunoblotting of ENSA in cortices from 6-month-old WT and Mme KO mice. Values indicated in the graph show the intensity of ENSA bands normalized to that of β-actin (n = 5 for each group). c, Aβ 42 ELISA of Tris-HCl-buffered saline-soluble fractions containing 1% Triton-X from hippocampi of WT mice after overexpression of active or inactive mutant NEP by the SFV gene expression system (n = 4 for each group). Results are expressed as the mean ±SEM. *P <0.05 (Student’s t -test).