Detection of Human Dopamine beta ‑Hydroxylase by Western Blot. Western blot shows lysates of human adrenal gland tissue. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human Dopamine beta ‑Hydroxylase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7376) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Dopamine beta ‑Hydroxylase at approximately 74-77 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
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: Dopamine beta-Hydroxylase
DBH (Dopamine beta -Hydroxylase) is a 77-78 kDa member of the Cu++ type II ascorbate-dependent monooxygenase family of enzymes. It is expressed in noradrenergic nerve terminals and adrenal medullary chromaffin cells, and serves as a catalyst for the conversion of dopamine into norepinepherine. Unlike its family member counterparts (TH and PNMT) that are cytosolic, DBH is embedded in the membranes of secretory vesicles. Human DBH is a 617 amino acid (aa) type II transmembrane glycoprotein (SwissProt # P09172). It contains an N-terminal 16 aa cytoplasmic region, plus a 580 aa lumenal domain (aa 38-617). The luminal region possesses one DOMON (dopamine beta -monooxygenase N-terminal) domain (aa 51-169) plus two consecutive monooxygenase motifs (aa 214-523). DBH exists as both a membrane-embedded 77-78 kDa isoform, and a soluble 73-75 kDa isoform. The latter may arise from cleavage following Gly39. Structurally, DBH will form a disulfide-linked homodimer, which then noncovalently associates with another DBH covalent homodimer, generating a functional 290 kDa homotetramer. Over aa 37‑617, human DBH shares 79% aa sequence identity with mouse DBH.
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