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Human Dopamine beta-Hydroxylase HEK293 Overexpression Lysate

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Specification

Cat. No.

NAE450

Neurotransmitter Associated Enzymes

Dopamine beta-Hydroxylase

Product Type

Lysates

Product Name

Human Dopamine beta-Hydroxylase HEK293 Overexpression Lysate

Size

300 µg

Price

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Product Description

This Human Dopamine beta-Hydroxylase overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Dopamine beta-Hydroxylase protein from the overexpression lysate was verified.

Expression Host

HEK293 Cells

Species

Human

Sequence Information

A DNA sequence encoding the human DBH (P09172) (Ser26-Gly603) was expressed with an N-terminal polyhistidine tag.

Molecule Mass

The recombinant human DBH comprises 598 amino acids and has a predicted molecular mass of 67.3 kDa. The apparent molecular mass of the protein is approximately 68 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Stability & Storage

Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.

Preparation Method

Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.

Lysis Buffer

Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.

Recommend Usage

1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.

Sample Buffer

1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

Synonyms

Human DBM Overexpression Lysate

Application

Western Blot (WB). Optimal dilutions/concentrations should be determined by the end user.

All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.