NKp30/NCR3 Overexpression Lysate

Name: NKp30/NCR3 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-13521

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-13521

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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 NKp30/NCR3 Overexpression Lysate

NKp30/NCR3 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC208631

Accession#: NM_147130

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

21.4 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 NKp30/NCR3 Overexpression Lysate

Western Blot: NKp30/NCR3 Overexpression Lysate [NBL1-13521]

Western Blot: NCR3 Overexpression Lysate (Adult Normal) [NBL1-13521] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NCR3.

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: NKp30/NCR3

NKp30, also referred to as natural cytotoxicity receptor 3 (NCR3), is a type I transmembrane receptor belonging to the immunoglobulin (Ig) superfamily that is expressed on resting and active natural killer (NK) cells with a role in triggering a cytotoxic response against tumor cells (1). The human NKp30 protein has six isoforms. The canonical isoform (NKp30a) is 177 amino acids (aa) in length with a theoretical molecular weight of 21.6 kDa (1,2). The NKp30 protein consists of a signal peptide, an Ig domain, a stalk region, a transmembrane domain, and an intracellular region (1-3). NKp30/NCR3 signaling relies on interaction with the immunoreceptor tyrosine-based activation motif (ITAM)-containing adaptor molecules CD3zeta and FcRgamma (1,4,5). Other NRC family members include NKp44 and NKp46 (1,3-6). NK cell activation and tumor surveillance occurs through receptor-ligand binding. Tumor-derived ligands for NKp30 include HLA-B-associated transcript 3 (BAT3)/Bcl2-associated anthogene 6 (BAG6) and B7-H6 (4-6). NKp30 has also been suggested to play a role in NK-dendritic cell (DC) crosstalk by promoting DC maturation and cytotoxicity of immature DCs, mediated in part by BAT3 (1,4,5).

Given its role in NK cell-mediated killing and activation, NKp30 is a potential target for cancer immunotherapies (6). For instance, Bi-specific T cell engagers (BiTE) is one immunotherapy approach targeting the NKp30/B7-H6 interaction where B7-H6-specific BiTE elicits an attack on tumor cells through interferon gamma (IFNgamma) production and T cell cytotoxicity (1). Another potential approach is NKp30-derived chimeric antigen receptors (CARs) where CAR-T cells that express chimeric NKp30 receptors can efficiently attack B7-H6 tumor cells (1,6).

References

1. Barrow, A. D., Martin, C. J., & Colonna, M. (2019). The natural cytotoxicity receptors in health and disease. Frontiers in Immunology, 10, 909. https://doi.org/10.3389/fimmu.2019.00909

2. Uniprot (O14931)

3. Luczo, J. M., Ronzulli, S. L., & Tompkins, S. M. (2021). Influenza A virus hemagglutinin and other pathogen glycoprotein interactions with NK cell natural cytotoxicity receptors NKp46, NKp44, and NKp30. Viruses, 13(2), 156. https://doi.org/10.3390/v13020156

4. Pazina, T., Shemesh, A., Brusilovsky, M., Porgador, A., & Campbell, K. S. (2017). Regulation of the Functions of natural cytotoxicity receptors by interactions with diverse ligands and alterations in splice variant expression. Frontiers in Immunology, 8, 369.

5. Kaifu, T., Escaliere, B., Gastinel, L. N., Vivier, E., & Baratin, M. (2011). B7-H6/NKp30 interaction: a mechanism of alerting NK cells against tumors. Cellular and Molecular Life Sciences : CMLS, 68(21), 3531-3539. https://doi.org/10.1007/s00018-011-0802-7

6. Pinheiro, P. F., Justino, G. C., & Marques, M. M. (2020). NKp30 - A prospective target for new cancer immunotherapy strategies. British Journal of Pharmacology, 177(20), 4563-4580. https://doi.org/10.1111/bph.15222

Alternate Names

CD337, NCR3

Gene Symbol

NCR3

Additional NKp30/NCR3 Products

Product Documents for NKp30/NCR3 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for NKp30/NCR3 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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