Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH17-05] to Human NKG2D antibody (Detector) (HA725300) and Recombinant Human NKG2D protein (HA211290) as the standard. The reference range value is 7.8-2,000 pg/mL.
ELISA
Use at an assay dependent concentration.
Target
Function
NKG2D is an activating receptor (transmembrane protein) belonging to the NKG2 family of C-type lectin-like receptors. NKG2D is encoded by KLRK1 (killer cell lectin like receptor K1) gene which is located in the NK-gene complex (NKC) situated on chromosome 6 in mice and chromosome 12 in humans. In mice, it is expressed by NK cells, NK1.1+ T cells, γδ T cells, activated CD8+ αβ T cells and activated macrophages. In humans, it is expressed by NK cells, γδ T cells and CD8+ αβ T cells. NKG2D recognizes induced-self proteins from MIC and RAET1/ULBP families which appear on the surface of stressed, malignant transformed, and infected cells.
Background References
1. Fuertes MB et al. Leveraging NKG2D Ligands in Immuno-Oncology. Front Immunol. 2021 Jul
2. Hartmann JA et al. Evasion of NKG2D-mediated cytotoxic immunity by sarbecoviruses. Cell. 2024 May
Sandwich ELISA analysis of human NKGD2(KLRK1) matched pair antibodies
Capture: HA725299, Human NKG2D Rabbit mAb [PSH17-04] Detector: HA725300, Human NKG2D Rabbit mAb [PSH17-05]
Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725299) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human NKG2D protein (HA211290) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA725300, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
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