TMPRSS2 Overexpression Lysate

Name: TMPRSS2 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-17121

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-17121

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for TMPRSS2 Overexpression Lysate

TMPRSS2 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC208677

Accession#: NM_005656

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.

Product Specifications

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.

TMW

53.9 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for TMPRSS2 Overexpression Lysate

Western Blot: TMPRSS2 Overexpression Lysate [NBL1-17121]

Western Blot: TMPRSS2 Overexpression Lysate [NBL1-17121] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -.TMPRSS2 Overexpression Lysate (approximate molecular weight of 53.9 KDa).

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: TMPRSS2

TMPRSS2, also called Epitheliasin in mice, is a 492 amino acid type II transmembrane serine protease located on human chromosome 21q22.3 that encodes at least 2 isoforms. This glycosylated serine protease (theoretical molecular weight 70kDa) is regulated by androgens and expressed on the plasma membrane of human bronchial epithelial cells, nasal goblet cells, small intestine epithelia, gastrointestinal tract, the stomach, kidneys and pancreas, with the greatest abundance found in the prostate gland. While the physiological function of TMPRSS2 remains unknown, the serine protease domain undergoes autocleavage and the 32 kDa domain is secreted enabling its interaction with the extracellular matrix. Fusions between TMPRSS2 and the ETS transcription factor genes ERG, ETV1, and ETV4 have been reported in prostate cancer with the TMPRSS2 ERG gene fusion resulting in ERG overexpression in 40-80% of these cases and often producing a more aggressive phenotype. Androgen signaling is disrupted in prostate cancers with the TMPRSS2 ERG fusion which contributes to the switch from androgen dependent to androgen independent prostate cancer (1,2).

TMPRSS2 has also been shown to play a critical role in the process of viral entry for coronaviruses and influenza viruses thru proteolytic activation of key viral glycoproteins. One pathway for SARS-CoV-2 infection, which causes the COVID-19 disease, involves binding of the SARS-CoV-2 Spike 'S' glycoprotein to the human ACE2 receptor and cleavage of S by TMPRSS2 at the cell surface to facilitate viral entry. TMPRSS2 mediates viral entry in a similar mechanism for other coronaviruses such as SARS-CoV and MERS. The broad-spectrum serine protease inhibitor, Camostat, is a TMPRSS2 inhibitor demonstrated to protect mice with lethal SARS-CoV infections (3).

References

1. Clark, J., Cooper, C. (2009) ETS gene fusions in prostate cancer. Nat Rev Urol 6, 429-439. PMID: 19657377

2. Duffy, MJ. (2014) Chapter One - PSA in Screening for Prostate Cancer: More Good than Harm or More Harm than Good? Adv Clin Chem. 66:1-23. PMID: 25344984

3. Shen LW, Mao HJ, Wu YL, Tanaka Y, Zhang W. (2017) TMPRSS2: A potential target for treatment of influenza virus and coronavirus infections. Biochimie. 142:1-10

Long Name

Transmembrane protease, serine 2

Alternate Names

Epitheliasin, PP9284, PRSS10

Gene Symbol

TMPRSS2

Additional TMPRSS2 Products

Product Documents for TMPRSS2 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for TMPRSS2 Overexpression Lysate

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.

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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