Detects human 14-3-3 zeta in direct ELISAs and Western blots. Detects Mouse and Rat 14-3-3 zeta in Western Blots. In direct ELISAs, no cross-reactivity with recombinant human 14-3-3 beta, theta, eta, gamma, sigma, or epsilon is observed.
Monoclonal Mouse IgG2B Clone # 818515
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human 14-3-3 zeta Asp2-Asn245 Accession # P63104
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Detection of Human, Mouse, and Rat 14‑3‑3 zeta by Western Blot.
Western blot shows lysates of SH‑SY5Y human neuroblastoma cell line, SK‑Mel‑28 human malignant melanoma cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and C6 rat glioma cell line. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human 14‑3‑3 zeta Monoclonal Antibody (Catalog # MAB2669) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for 14‑3‑3 zeta at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
14‑3‑3 zeta was detected in immersion fixed paraffin-embedded sections of human squamous cell carcinoma using Mouse Anti-Human 14‑3‑3 zeta Monoclonal Antibody (Catalog # MAB2669) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei and plasma membrane. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human 14‑3‑3 zeta by Simple WesternTM.
Simple Western lane view shows lysates of SH‑SY5Y human neuroblastoma cell line, loaded at 0.5 mg/mL. A specific band was detected for 14‑3‑3 zeta at approximately 34 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human 14‑3‑3 zeta Monoclonal Antibody (Catalog # MAB2669). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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 (gene name YWHAZ) is a 245 amino acid, 27 kDa protein.
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