Detection of Human/Mouse 14‑3‑3 zeta by Western Blot.
Western blot shows lysates of SH‑SY5Y human neuroblastoma cell line, Raji human Burkitt's lymphoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, and PC‑12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.25 µg/mL Goat Anti-Human/ Mouse 14‑3‑3 zeta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2669) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). For additional reference, recombinant human 14-3-3 beta, epsilon, eta, gamma, sigma, and zeta (2 ng/lane) were included. A specific band for 14-3-3 zeta was detected at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: 14-3-3 zeta
14-3-3 proteins are a highly conserved family of homo- and heterodimeric phosphoserine/threonine-binding proteins present in high abundance in all eukaryotic cells. 14-3-3 proteins were the first polypeptides shown to have pSer/Thr binding properties, generally recognizing the consensus sequences RSXpSXP and RXY/FXpSXP (where X is any amino acid). 14-3-3 proteins act as key regulators of intracellular signal transduction through their ability to bind specific motifs phosphorylated on serine or threonine. For example, the binding of 14-3-3 to phosphorylated BAD blocks its proapoptotic association with Bcl-XL. There are at least seven distinct 14-3-3 genes in vertebrates, alpha/beta, epsilon, eta, gamma, theta, sigma and zeta ( alpha / beta, epsilon, eta, gamma, tau / theta, sigma, and zeta /δ). 14-3-3 zeta, also known as Tyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase Activation Protein, zeta isoform (YWHAZ) is a 245 amino acid, 27 kDa protein.
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