NKG2A/CD159a/KLRC1 Overexpression Lysate

Name: NKG2A/CD159a/KLRC1 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-12361

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-12361

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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 NKG2A/CD159a/KLRC1 Overexpression Lysate

NKG2A/CD159a/KLRC1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC211621

Accession#: NM_213657

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

24 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 NKG2A/CD159a/KLRC1 Overexpression Lysate

Western Blot: NKG2A/CD159a/KLRC1 Overexpression Lysate [NBL1-12361]

Western Blot: KLRC1 Overexpression Lysate (Adult Normal) [NBL1-12361] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for KLRC1.

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: NKG2A/CD159a

NK group 2 member A (NKG2A), also known as killer lectin like receptor c1 (KLRC1) and CD159a, is a ~38 kDa type II transmembrane protein belonging to the C-type lectin superfamily and plays a role in suppression of cytotoxic CD8+ T cell activity and functions as an immune checkpoint (1,2). NKG2A belongs to a group of natural killer (NK) cell inhibitor receptors (IRs) which includes killer immunoglobulin receptors (KIRs), lymphocyte activation gene-3 (LAG-3), T cell immunoreceptor with Ig and ITIM domains (TIGIT), programmed death ligand-1 (PD-1), and T cell immunoglobulin mucin-3 (TIM-3) (1,3). NKG2A is synthesized as a protein of 233 amino acids (aa) with a theoretical molecular weight of 26.3 kDa and is expressed on CD56hi NK cells, NKT cells, and some CD8+ alphabeta T cells (2,4). Together with CD94, NKG2A forms a heterodimer and acts as a receptor for the MHC Class I molecular human leukocyte antigen (HLA)-E (1-3). The NKG2A/CD94 receptor interaction with the HLA-E ligand results in phosphorylation of NKG2A's immunoreceptor tyrosine-based inhibition motifs (ITIMs), causing suppression of activating signals from immunoreceptor tyrosine-based activation motif (ITAM)- containing T cell receptors (TCR) or other activating receptors like NKG2D, via the intracellular phosphatase SHP1 (2,3,5). Overall, cancer cells utilize the NKG2A-HLA-E interaction and signaling to evade immune surveillance by NK cells and T cells (1-3,5-6).

Similar to cytotoxic T lymphocyte associated protein 4 (CTLA-4) and PD-1, NKG2A has become a prominent target for immune checkpoint blockade and cancer immunotherapy (1-3,5-6). Monalizumab is a novel IgG4 monoclonal antibody developed for blocking NKG2A's interaction with HLA-E and has shown promising results in clinical trials (1-3,5-6). In addition to being used as a single agent, it is being studied as a possible combination therapy with other blocking antibodies such as anti-PD-L1 and anti-epidermal growth factor receptor (EGFR) (2,5,6).

References

1. Alfarra, H., Weir, J., Grieve, S., & Reiman, T. (2020). Targeting NK Cell Inhibitory Receptors for Precision Multiple Myeloma Immunotherapy. Frontiers in immunology, 11, 575609. https://doi.org/10.3389/fimmu.2020.575609

2. Creelan, B. C., & Antonia, S. J. (2019). The NKG2A immune checkpoint - a new direction in cancer immunotherapy. Nature reviews. Clinical oncology, 16(5), 277-278. https://doi.org/10.1038/s41571-019-0182-8

3. Zaghi, E., Calvi, M., Marcenaro, E., Mavilio, D., & Di Vito, C. (2019). Targeting NKG2A to elucidate natural killer cell ontogenesis and to develop novel immune-therapeutic strategies in cancer therapy. Journal of leukocyte biology, 105(6), 1243-1251. https://doi.org/10.1002/JLB.MR0718-300R

4. Uniprot (P26715)

5. Borst, L., van der Burg, S. H., & van Hall, T. (2020). The NKG2A-HLA-E Axis as a Novel Checkpoint in the Tumor Microenvironment. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(21), 5549-5556. https://doi.org/10.1158/1078-0432.CCR-19-2095

6. Andre, P., Denis, C., Soulas, C., Bourbon-Caillet, C., Lopez, J., Arnoux, T., Blery, M., Bonnafous, C., Gauthier, L., Morel, A., Rossi, B., Remark, R., Breso, V., Bonnet, E., Habif, G., Guia, S., Lalanne, A. I., Hoffmann, C., Lantz, O., Fayette, J., ... Vivier, E. (2018). Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells. Cell, 175(7), 1731-1743.e13. https://doi.org/10.1016/j.cell.2018.10.014

Long Name

Natural Killer G2A

Alternate Names

CD159a, Klrc1, NKG2

Gene Symbol

KLRC1

Additional NKG2A/CD159a Products

Product Documents for NKG2A/CD159a/KLRC1 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for NKG2A/CD159a/KLRC1 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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