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Human COMT Baculovirus-Insect Overexpression Lysate

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Specification

Cat. No.

NAE448

Neurotransmitter Associated Enzymes

COMT

Product Type

Lysates

Product Name

Human COMT Baculovirus-Insect Overexpression Lysate

Size

300 µg

Price

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Product Description

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

Expression Host

Baculovirus-Insect Cells

Species

Human

Sequence Information

A DNA sequence encoding the human COMT (NP_000745.1) (Met1-Pro271) was expressed with a polyhistidine tag at the C-terminus.

Molecule Mass

The recombinant human COMT consists of 282 amino acids and predicts a molecular mass of 31.5 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.

Preparation Method

Cell lysate was prepared by homogenization 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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.

Sample Buffer

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

Synonyms

Human HEL-S-98n 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.