Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH01-26] to Human Mesothelin antibody (Capture) (HA723287) and Recombinant Human Mesothelin protein (HA210521) as the standard. The reference range value is 937.5-30,000 pg/ml.
ELISA
Use at an assay dependent concentration.
Target
Function
Mesothelin, also known as MSLN, is a protein that in humans is encoded by the MSLN gene. Mesothelin is a 40 kDa protein that is expressed in mesothelial cells. The protein was first identified by its reactivity with monoclonal antibody K1. Subsequent cloning studies showed that the mesothelin gene encodes a precursor protein that is processed to yield mesothelin which is attached to the cell membrane by a glycophosphatidylinositol linkage and a 31-kDa shed fragment named megakaryocyte-potentiating factor (MPF). Although it has been proposed that mesothelin may be involved in cell adhesion, its biological function is not known. A knockout mouse line that lacks mesothelin reproduces and develops normally. Mesothelin is over expressed in several human tumors, including mesothelioma, ovarian cancer, pancreatic adenocarcinoma, lung adenocarcinoma, and cholangiocarcinoma. Mesothelin binds MUC16 (also known as CA125), indicating that the interaction of mesothelin and MUC16 may contribute to the implantation and peritoneal spread of tumors by cell adhesion. The region (residues 296-359) consisting of 64 amino acids at the N-terminus of cell surface mesothelin has been identified as the functional binding domain (named IAB) for MUC16/CA125, suggesting the mechanism of mesothelin acting as a MUC16/CA125 functional partner in cancer development.
Background References
1. Klampatsa A et al. Mesothelin-targeted CAR-T cell therapy for solid tumors. Expert Opin Biol Ther. 2021 Apr
2. Faust JR et al. Mesothelin: An Immunotherapeutic Target beyond Solid Tumors. Cancers (Basel). 2022 Mar
Sandwich ELISA analysis of Human Mesothelin matched pair antibodies
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723287) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Mesothelin protein (HA210521) starting from 30,000 pg/ml to 0 pg/ml and detect antibody (HA723289H, HRP, 0.2 µg/ml) for 1 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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