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CD73 Cell Lysate | NT5E cell lysate

Cynomolgus CD73 HEK293 Overexpression Lysate

Applications
Western Blot
Synonyms
CD73; Cynomolgus CD73 HEK293 Overexpression Lysate; Cynomolgus CD73/NT5E HEK293 Cell Lysate (WB positive control); 5'-nucleotidase; ecto (CD73); NT5E cell lysate
Ordering
For Research Use Only!
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading 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.
Sequence Positions
532aa
Sequence
Met1-Lys547
Applicable Applications for NT5E cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Cynomolgus
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 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the cynomolgus NT5E (EHH53214.1) (Met1-Lys547) was expressed with a polyhistidine tag at the C-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. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for NT5E cell lysate
5'-nucleotidase, also known as NT5E, NTE, and CD73, is a cell membrane protein that belongs to the 5'-nucleotidase family. CD73 is a glycosylphosphatidylinositol (GPI) anchored purine salvage enzyme expressed on the surface of human T and B lymphocytes. CD73 catalyzes the conversion of purine and pyrimidine ribo-and deoxyribonucleoside monophosphates to the corresponding nucleosides. CD73 serves as a costimulatory molecule in activating T cells. CD73 generated adenosine functions in cell signaling in many physiologic systems, including intestinal epithelium, ischemic myocardium, and cholinergic synapses. CD73 might mediate lymphocyte-stromal cell interactions or condition the local microenvironment to facilitate lymphocyte development and/or function. In CD73-depleted cells, surface levels of the leukocyte adhesion molecules ICAM-1, VCAM-1, and E-selectin increase. CD73 produces extracellular adenosine, which then acts on G protein-coupled purinergic receptors to induce cellular responses. CD73 has also been reported to regulate the expression of pro-inflammatory molecules in mouse endothelium.

This Cynomolgus CD73 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD73 protein from the overexpression lysate was verified.
Product Categories/Family for NT5E cell lysate
References
Resta R. et al., 1997, Cell Signal. 9 (2): 131-9. Yamashita Y. et al., 1998, Eur J Immunol. 28 (10): 2981-90. Louis NA. et al., 2008, J Immunol. 180 (6): 4246-55. Grunewald JK. et al., 2010, J Inflamm. 7 (1): 10.

NCBI and Uniprot Product Information

Molecular Weight
The recombinant cynomolgus NT5E comprises 532 amino acids and has a calculated molecular mass of 59.2kDa.
The apparent molecular mass of it is approximately 59kDa respectively in SDS-PAGE.

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Product Notes

The NT5E (Catalog #AAA8117887) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 532aa. AAA Biotech's CD73 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the NT5E for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Lys54 7. It is sometimes possible for the material contained within the vial of "CD73, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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