Human ENTPD2 Baculovirus-Insect cells Overexpression Lysate

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Human ENTPD2 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human ENTPD2 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of ENTPD2 protein (Cat: 11734-H07B1) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the mature form of human ENTPD2 (Q9Y5L3) (Thr29-Asp460) was expressed, with a polyhistidine tag at the N-terminus.
Molecule Mass
The recombinant human human ENTPD2 consists of 448 amino acids and predicts a molecular mass of 49.3 KDa. It migrates as an approximately 59 KDa band in SDS-PAGE under reducing conditions.

Human ENTPD2 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 ENTPD2 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human CD39L1 Overexpression Lysate; Human NTPDase-2 Overexpression Lysate; Human RP11-229P13.11-001 Overexpression Lysate

ENTPD2 Background Information

NTPDase 2, also known as ENTPD2, belongs to the ecto-nucleoside triphosphate diphosphohydrolase family (E-NTPDase). Members of E-NTPDase family are nucleotidases able to hydrolyze 5′-nucleoside tri- and/or diphosphates; the main role of these enzymes is the termination of purinergic signaling. NTPDases are ubiquitous and were previously shown in other parasites including the trypanosomatides of genus Leishmania and in T. brucei. NTPase activity would act as a timer and is crucial to T. gondii infection. In L. pneumophila it was demonstrated that an E-NTPDase, similar to CD39, is essential for intracellular bacterial multiplication. NTPDase 2 is an integral membrane protein. In the nervous system, it could hydrolyze ATP and other nucleotides to regulate purinergic neurotransmission. Alternative splicing of NTPDase 2 gene results in multiple transcript variants.
Full Name
ectonucleoside triphosphate diphosphohydrolase 2
References
  • Harden TK, et al. (2005) Regulation of P2Y1 receptor-mediated signaling by the ectonucleoside triphosphate diphosphohydrolase isozymes NTPDase1 and NTPDase2. Mol Pharmacol. 67(1): 114-22.
  • Dranoff JA, et al. (2005) Ectonucleotidase NTPDase2 is selectively down-regulated in biliary cirrhosis. J Investig Med. 52(7):475-82.
  • Jhandier MN, et al.. (2005) Portal fibroblasts regulate the proliferation of bile duct epithelia via expression of NTPDase2. J Biol Chem. 280(24):22986-92.
  • Mukasa T, et al. (2005) Either the carboxyl- or the amino-terminal region of the human ecto-ATPase (E-NTPDase 2) confers detergent and temperature sensitivity to the chicken ecto-ATP-diphosphohydrolase (E-NTPDase 8). Biochemistry. 44(33):11160-70.
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