NKp46/NCR1 Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-13519
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for NKp46/NCR1 Overexpression Lysate
NKp46/NCR1 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC208943
Accession#: NM_004829
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.
Expression Host: HEK293T
Plasmid: RC208943
Accession#: NM_004829
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.
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
34.3 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.
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 NKp46/NCR1 Overexpression Lysate
![Western Blot: NKp46/NCR1 Overexpression Lysate [NBL1-13519]](https://resources.bio-techne.com/images/products/NCR1-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-13519-img0002.jpg "Western Blot: NKp46/NCR1 Overexpression Lysate [NBL1-13519]")
Western Blot: NKp46/NCR1 Overexpression Lysate [NBL1-13519]
Western Blot: NCR1 Overexpression Lysate (Adult Normal) [NBL1-13519] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NCR1.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: NKp46/NCR1
NKp46 plays a critical role in the NK-cell lysis of allogenic, autologous, and xenogeneic cells (3). Activation of NKp46 signaling is mediated via its association with immunoreceptor tyrosine-based activation motif (ITAM) bearing molecules CD3zeta and FcRgamma (3). Upon receptor-engagement NK-cell cytotoxic activity and cytokine release occurs (3). This NKp46 signaling can be blocked with anti-NKp46 monoclonal antibody mediated cross-linking which causes calcium release and interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) secretion (3). NKp46 recognizes multiple ligands presented on tumor cells or pathogens and infected cells including influenza virus hemagglutinin and the parainfluenza virus hemagglutinin neuraminidase (2). Additional ligands for the NKp46 receptor include heparan sulfate (HS) glycosaminoglycans (GAGs), hemagglutinin of human vaccina virus, and vimentin expressed on cells infected with Mycobacterium tuberculosis (3).
NKp46 and its NCR counterparts have shown to be successful targets for cancer immunotherapy. More specifically, human and mouse NKp46 has shown promise in virotherapy due to its ability to stimulate NK cell activation by binding to reovirus sigma1 protein in a sialic acid-dependent manner (3). Another approach for NKp46 in cancer immunotherapy is through the use of NCR based chimeric antigen receptors (CARs). In this therapy, NCR1 supplied human T cells with anti-tumor properties via cytokine secretion, upregulation of activation markers, improved expansion, and cytotoxicity in both mouse and in vitro models (3).
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. Mandelboim, O., & Porgador, A. (2001). NKp46. The international journal of biochemistry & cell biology, 33(12), 1147-1150. https://doi.org/10.1016/S1357-2725(01)00078-4
3. 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
Alternate Names
CD335, Ly94, MAR-1, NCR1
Gene Symbol
NCR1
Additional NKp46/NCR1 Products
Product Documents for NKp46/NCR1 Overexpression Lysate
Product Specific Notices for NKp46/NCR1 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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