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Human DCLK1 Baculovirus-Insect cells Overexpression Lysate

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Specification

Cat. No.

NRT996

Neurotransmitter Receptors, Transporters, and Ion Channels

DCLK1

Product Type

Lysates

Product Name

Human DCLK1 Baculovirus-Insect cells Overexpression Lysate

Size

300 µg

Price

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Synonyms

Human CL1 Overexpression Lysate; Human CLICK1 Overexpression Lysate; Human DCAMKL1 Overexpression Lysate; Human DCDC3A Overexpression Lysate; Human DCLK Overexpression Lysate

Description

This Human DCLK1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control.

Species

Human

Molecule Mass

The secreted recombinant human DCLK1 consists of 707 amino acids and predicts a molecular mass of 78.5 KDa. The apparent molecular mass of the protein is approximately 64 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.

Application

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

Expression Host

Baculovirus-Insect cells

Sequence Information

A DNA sequence encoding the human DCLK1 ( O15075-1) (Met 1-Val 705) was expressed and purified with two additional amino acids (Gly & Pro ) 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).

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