Biolegend, 716755, Recombinant Human KIR3DL3 (carrier free), 50μg

Killer cell immunoglobulin-like receptor (KIR) genes coding for the KIR receptors are in a 100-200 Kb region of the Leukocyte Receptor Complex (LRC). Located on chromosome 19q13.4, KIR genes codes for receptors expressed by the Natural Killer cells. To date, 16 human KIR genes have been identified. Their nomenclature depends on the number of domains of extracellular immunoglobulins (D), the length of the intra cytoplasmic tail will be either long (L), or short (S). From the KIR inhibitory receptor family, KIR3DL3 is a unique member of this family with only one immunoreceptor tyrosine-based inhibitory motif (ITIM). KIR3DL3 encodes at least 93 amino acid sequences and all encoded amino acid sequences are full-length proteins. KIR3DL3 is expressed in CD56 dim NK and terminally differentiated effector memory CD8+ T cells. However, it is rarely expressed in normal tissues. KIR3DL3 may be involved in the formation of the placenta. KIR3DL3, is an inhibitory receptor for B7- H7/HHLA2. B7-H7-mediated signaling via KIR3DL3 inhibit T-cell activation and NK cell cytolytic activity. B7-H7 is involved in the aggregation of KIR3DL3 at the immune synapse. After being recruited, ITIM on KIR3DL3 is phosphorylated to recruit Src homology 2-containing protein-tyrosine phosphatases 1 and 2 (SHP1/2). The KIR3DL3 and B7-H7 interaction suppresses T cells and NK cells and reduces cytokine production, including interferon-gamma (IFNγ), TNF-α, IL-5, IL-10, IL-13, IL-17A and IL-22. It has been proposed that an immunotherapy that combines KIR3DL3 and PD-1, may show better antitumor efficacy.
50μg
Source: Human KIR3DL3, amino acids Gln26-Leu322 (Accession # Q8N743), with a C-terminal 8xHis tag was expressed in CHO cells.
Molecular Mass: The 308 amino acid recombinant protein has a predicted molecular mass of approximately 33.6 kD. The protein migrates approximately at 40-50 kD in SDS-PAGE in DTT-reduced and non-reduced conditions.
N-terminalSequence Analysis: The predicted N-terminal amino acid is Gln.
Purity: > 95%, as determined by Coomassie stained SDS-PAGE.
Formulation: Lyophilized from 0.22 µm filtered protein solution in PBS, pH 6.5 with 5% trehalose.
Endotoxin Level: Less than 0.1 EU per μg of protein as determined by the LAL method
Concentration: 50 µg size lyophilized
Storage & Handling: Unopened vial can be stored between 2°C -4°C for up to two weeks and at -20°C or colder until the expiration date. Reconstitute lyophilized protein in sterile water preferably in sterile deionized water. Before reconstitution, make sure sterile water and the product are at room temperature. Quickly spin the vial or gently tap down on the vial to make sure the lyophilized product is at the bottom of the vial before opening. Use aseptic techniques to add the required volume of reconstitution buffer (sterile water) to the vial, to obtain the recommended stock concentration 250 μg/mL. Close the vial and leave at an ambient temperature for 15-20 minutes. Then gently invert the vial several times or until all the lyophilized product dissolves. Leave the vial at room temperature for another 15 minutes. If small particulates are still observed after 15 minutes, incubate at room temperature for an additional 30 minutes and leave the vial at 2°C - 8°C overnight. Next day, invert the vial several times or gently pipette the solution up and down before use. If needed, transfer the reconstituted stock solution to a sterile container for additional dilution to no less than 100 μg/mL. Small working aliquots in polypropylene tubes can be made after reconstitution and store the vials at -20°C or lower. Avoid freeze/thaw cycles. Carrier protein such as 0.2 - 1% endotoxin-free BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to two weeks or stored at -20°C or colder for up to 3 months.
Activity: Immobilized human KIR3DL3 binds to human B7-H7 in a dose-dependent manner. ED50 for this effect is 0.08-0.56 µg/mL.
Application: Bioassay
Application Notes: Our lyophilized proteins are validated in-house to maintain activity after shipping at ambient temperature and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
Structure: Monomer
Distribution: CD56dim NK and terminally differentiated effector memory CD8+ T cells
Function: inhibits T-cell activation and NK cell cytolytic activity
Interaction: T-cells and NK cells
Ligand/Receptor: B7-H7/HHLA2
Bioactivity: Immobilized human KIR3DL3 binds to human B7-H7 in a dose-dependent manner. ED50 for this effect is 0.08-0.56 µg/mL.
Cell Type: Lymphocytes, NK cells, T cells
Biology Area: Cell Biology, Immunology, Innate Immunity
Molecular Family: CD Molecules
Antigen References: Trowsdale J, 2001. Immunity. 15:363-374. Kulkarni S, et al. 2008. Semin Immunol. 20:343-352. Marsh SGE, et al. 2002. Immunogenetics. 55:220-226. Vilches C. and Parham P. 2002. Annu Rev Immunol. 20:217-251. Robinson J, et al. 2013. Nucleic Acids Res. 41:1234-1240. González-Galarza FF, et al. 2015. Nucleic Acids Res. 43:D784-D788. K.S. Campbell. 2021. Cancer Immunol. Res. 9:128. Bhatt RS, et al. 2021. Cancer Immunol Res. 9:156-169. Pende D, et al. 2019. Front. Immunol. 10:1179. L.A. Leaton, et al. 2019. Front. Immunol. 10, p.24. Wei Y, et al. 2021. Sci. Immunol. 6:61. Bolandi N, et al. 2021. Int. J. Mol. Sci. 22:10719.
Gene ID: 115653
UniProt: View information about KIR3DL3 on UniProt.org
Regulatory Status: RUO
Other Names: Killer cell immunoglobulin-like receptor 3DL3, CD158 antigen-like family member Z, Killer cell inhibitory receptor 1, CD158z
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A: • Each lot of product is quality-tested for bioactivity as indicated on the data sheet.
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Q: How does the activity of your recombinant proteins compare to competitors?
A: We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
Q: What is the specific activity or ED50 of my recombinant protein?
A: The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.
Q: Have your recombinants been tested for stability?
A: Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
Q: Does specific activity of a recombinant protein vary between lots?
A: Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
Q: How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
A: Use formula Specific activity (Units/mg) = 10^6/ ED50 (ng/mL)