Human DCLK1 Baculovirus-Insect cells Overexpression Lysate

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Human DCLK1 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human DCLK1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of DCLK1 protein (Cat: 11588-H20B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human DCLK1 (O15075-1) (Met 1-Val 705) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Molecule Mass
The recombinant human DCLK1/GST chimera consists of 942 amino acids and has a calculated molecular mass of 106 KDa. It migrates as an approximately 105 KDa band as predicted in SDS-PAGE under reducing conditions.

Human DCLK1 Baculovirus-Insect cells Overexpression Lysate: Usage Guide

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).
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.

Human DCLK1 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

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

DCLK1 Background Information

DCAMKL1, also known as DCLK1, is a member of the protein kinase superfamily and the doublecortin family. It contains two N-terminal doublecortin domains, which bind microtubules and regulate microtubule polymerization, a C-terminal serine/threonine protein kinase domain, which shows substantial homology to Ca2+/calmodulin-dependent protein kinase, and a serine/proline-rich domain in between the doublecortin and the protein kinase domains, which mediates multiple protein-protein interactions. DCAMKL1 is involved in several different cellular processes, including neuronal migration, retrograde transport, neuronal apoptosis and neurogenesis. Its microtubule-polymerizing activity is independent of its protein kinase activity. DCAMKL1 may be involved in a calcium-signaling pathway controlling neuronal migration in the developing brain. It may also participate in functions of the mature nervous system.
Full Name
doublecortin-like kinase 1
References
  • Sossey-Alaoui K, et al. (1999) DCAMKL1, a brain-specific transmembrane protein on 13q12.3 that is similar to doublecortin (DCX). Genomics. 56 (1): 121-6.
  • Matsumoto N, et al. (1999) Genomic structure, chromosomal mapping, and expression pattern of human DCAMKL1 (KIAA0369), a homologue of DCX (XLIS). Genomics. 56 (2): 179-83.
  • Lin PT, et al. (2001) DCAMKL1 encodes a protein kinase with homology to doublecortin that regulates microtubule polymerization. J Neurosci. 20 (24): 9152-61.
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