| Species : |
Mouse |
| Source : |
E.coli |
| Tag : |
GST&His |
| Protein Length : |
26-106 aa |
| Form : |
Lyophilized from sterile 20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300. |
| Bio-activity : |
Nucb2 (Nucleobindin-2), a calcium-binding protein, is further cleaved into NES1 (Nesfatin-1). NES1 is an anorexigenic peptide and seems to participate in hypothalamic pathways regulating food intake and energy homeostasis, acting in a leptin-independent manner. GADD45A (Growth arrest and DNA damage-inducible protein GADD45 alpha) has been identified as an interactor of Nucb2. Besides, rat GADD45A shares similarities with mouse NES1 in amino acid sequence with the identity of 98%. Thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse NES1 and recombinant rat GADD45A. Briefly, NES1 were diluted serially in PBS, with 0.01%BSA (pH 7.4). Duplicate samples of 100uL NES1 were then transferred to GADD45A-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-NES1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37°C. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of NES1 and GADD45A was shown, and this effect was in a dose dependent manner. |
| Molecular Mass : |
39.5kDa |
| Purity : |
> 97% as determined by SDS-PAGE. |
| Storage : |
Avoid repeated freeze/thaw cycles. Store at -20°C for 12 months. Aliquot and store at -80°C for 12 months. |
| Reconstitution : |
Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex. |
