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Human DCAMKL1 / DCLK1 Insect Cell Lysate (WB positive control)

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Human DCLK1 Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Baculovirus-Insect cells
Product Description:Baculovirus-Insect Cell lysate that Human DCAMKL1 / DCLK1 transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
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.
Predicted N Terminal:Gly
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.
Human DCLK1 Transfected / Overexpression Cell Lysate Usage Guide
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.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
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℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
DCAMKL1/DCLK1 Background

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.

Human DCAMKL1/DCLK1 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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    Catalog: 11588-HNCBL-300
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