CD160 Overexpression Lysate

Name: CD160 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-08908

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-08908

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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 CD160 Overexpression Lysate

CD160 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC204886

Accession#: NM_007053

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

19.6 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 CD160 Overexpression Lysate

Western Blot: CD160 Overexpression Lysate [NBL1-08908]

Western Blot: CD160 Overexpression Lysate (Adult Normal) [NBL1-08908] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD160.

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: CD160

CD160, also known as BY55 and NK1, belongs to a class of natural killer (NK) receptors known as the major histocompatibility complex (MCH) class I-dependent NK cell activating receptor (1). CD160 exists as either a glycosylphosphatidylinositol (GPI)-anchored isoform or a transmembrane protein and is expressed on NK cells and cytotoxic T lymphocytes, as well as endothelial cells (1-3). CD160 has both stimulatory and inhibitory receptor functions and regulates cell differentiation and activation (2). The human CD160 protein is 181 amino acids (aa) in length with a theoretical molecular weight (MW) of 19.8 kDa (2,3). However, given that the CD160 protein is subject to glycosylation, addition of disulfide bonds, lipidation, and propitiation, the observed MW is often much higher (2). The human CD160 protein consists of a single immunoglobulin (Ig)-like domain (25-133 aa) and does not contain any immunoreceptor tyrosine-based activation motifs (ITSMs) (2). Thus, CD160 requires the recruitment of adaptor proteins such as phosphoinositide-3 kinase (PI3K) to promote cytotoxic functions including cytokine secretion (1,3).

Identified ligands for CD160 include MHC class I proteins and herpes virus entry mediators (HVEM), each of which has a different effect on NK or T cells (1,3-5). CD160 binds to MHC class I proteins with weaker affinity, but the interaction causes NK cell cytotoxic function and cytokine production (1,3). CD160 engagement of MHC class I proteins and/or HVEM within NK cells promotes ERK1/2 and AKT activation and production of interferon gamma (IFNgamma) (5). Conversely, CD160 binding of HVEM in CD4+ T cells induces inhibitory signaling, signifying both a stimulatory and inhibitory role for CD160 as well as cell-specific context (4,5). Additionally, CD160 plays a role in diseases including certain types of cancer, such as thyroid cancer and colon cancer, viral infections, and autoimmune diseases (3,5). CD160 expression levels on CD8+ T cells have been shown to be elevated in chronic viral infections such as human immunodeficiency virus (HIV) and Epstein Barr virus (EBV) (3). In addition, when CD160 is co-expressed with the programmed cell death protein 1 (PD-1) receptor, T cell exhaustion occurs due to persistent stimulation, leading to inhibited immune response and ability to combat infection (3). CD160 may be a promising therapeutic target in cancer as murine studies have demonstrated that blocking the CD160-HVEM pathway causes a regression of neoplastic tumors in a thyroid cancer model (3-5).

References

1. Le Bouteiller P, Tabiasco J, Polgar B, et al. CD160: a unique activating NK cell receptor. Immunol Lett. 2011;138(2):93-96. https://doi.org/10.1016/j.imlet.2011.02.003

2. Uniprotkb (O95971)

3. Piotrowska M, Spodzieja M, Kuncewicz K, Rodziewicz-Motowidlo S, Orlikowska M. CD160 protein as a new therapeutic target in a battle against autoimmune, infectious and lifestyle diseases. Analysis of the structure, interactions and functions. Eur J Med Chem. 2021;224:113694. https://doi.org/10.1016/j.ejmech.2021.113694

4. Cai G, Freeman GJ. The CD160, BTLA, LIGHT/HVEM pathway: a bidirectional switch regulating T-cell activation. Immunol Rev. 2009;229(1):244-258. https://doi.org/10.1111/j.1600-065X.2009.00783.x

5. Sedy JR, Ramezani-Rad P. HVEM network signaling in cancer. Adv Cancer Res. 2019;142:145-186. https://doi.org/10.1016/bs.acr.2019.01.004

Alternate Names

BY55, CD160, NK1, NK28

Gene Symbol

CD160

Additional CD160 Products

Product Documents for CD160 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for CD160 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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