TMPRSS3 Overexpression Lysate Summary
Description |
TMPRSS3 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC224957
Accession#: NM_032401
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
Gene |
TMPRSS3 |
Applications/Dilutions
Dilutions |
|
Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for TMPRSS3 Overexpression Lysate
Background
Extracellular proteases mediate the digestion of neighboring extracellular matrix components in initial tumor growth, allow desquamation of tumor cells into the surrounding environment, provide the basis for invasion of basement membranes in targeted metastatic organs and are required for release and activation of many growth and angiogenic factors. The TMPRSS3 (also known as ECHOS1) gene, which encodes a transmembrane serine protease, has been found to be responsible for two non-syndromic recessive deafness loci located on human chromosome 21q22.3, DFNB8 and DFNB10.TMPRSS3, a 437 amno acid membrane bound serine protease and a member of the S1 peptidase family. TMPRSS3 contains an amino-terminal signalanchor sequence and a glycosylated extracellular region containing the serine protease domain. Two novel missense mutations of TMPRSS3, W251C and P404L, alter the highly conserved amino acids of the serine protease domain. TMPRSS3 is expressed in many tissues, including fetal cochlea, a subset of pancreatic cancer and various other cancer tissues. TMPRSS3 is also overexpressed in cancer, suggesting that it may be important for processes in metastasis formation and tumor invasion.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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