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Human Tyrosine Hydroxylase Baculovirus-Insect cells Overexpression Lysate

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Specification

Cat. No.

NSC715

Category

Neural Stem Cell Growth Factors

Product Type

Lysates

Product Name

Human Tyrosine Hydroxylase Baculovirus-Insect cells Overexpression Lysate

Size

300 µg

Price

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Synonyms

Human DYT14 Overexpression Lysate; Human DYT5b Overexpression Lysate; Human TYH Overexpression Lysate

Species

Human

Molecule Mass

The recombinant human TH consists of 515 amino acids and has a calculated molecular mass of 57.6 kDa.

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.

Product Description

This Human Tyrosine Hydroxylase overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Tyrosine Hydroxylase protein (Cat: 10684-H07B) from the overexpression lysate was verified.

Expression Host

Baculovirus-Insect cells

Sequence Information

A DNA sequence encoding the human TH isoform 2 (P07101-3) (Pro 2-Gly 497) was fused with a polyhistidine tag at the N-terminus.

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).

Application

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

Related Pathways

G Protein-coupled Receptors Signaling

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