Detection of Human Mesothelin by Simple WesternTM.
Simple Western lane view shows lysates of human ovarian cancer tissue and human lung tissue, loaded at 0.2 mg/mL. A specific band was detected for Mesothelin at approximately 65 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human Mesothelin Monoclonal Antibody (Catalog # MAB32653). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
*Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Detection of Human Mesothelin by Western Blot. Western blot shows lysates of human ovarian cancer tissue. PVDF Membrane was probed with 2 µg/mL of Human Mesothelin Monoclonal Antibody (Catalog # MAB32653) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Mesothelin at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Mesothelin, also known as CAK1 and ERC, is derived from a 70 kDa precursor that also includes Megakaryocyte Potentiating Factor (MPF) (1-3). The 70 kDa precursor is expressed on the cell surface where it is cleaved at a dibasic proteolytic site to release the 32 kDa glycosylated MPF (3, 4). MPF is a cytokine that potentiates IL-3 induced megakaryocyte colony formation (2, 5). The term Mesothelin refers to the 40 kDa glycosylated protein which remains attached to the cell surface via a GPI linkage. Alternate splicing generates additional Mesothelin isoforms that have either an eight amino acid insertion following Ser408 or a substituted C-terminal region with no GPI anchor (6). Within aa 29-580, human Mesothelin shares 59% sequence identity with mouse and rat Mesothelin. Mesothelin is normally expressed on mesothelial cells in the pleura, pericardium, and peritoneum as well as in the developing and postnatal pancreas (1, 7). It is upregulated in mesotheliomas and a range of carcinomas and adenomas (8-11). Mesothelin promotes tumor cell proliferation, migration, anchorage-independent growth, and tumor progression (10, 12). It is coexpressed with the tumor antigen CA125/MUC16 on advanced ovarian adenocarcinomas and interacts with this molecule to support cell adhesion (13). A soluble form of Mesothelin is released from tumor cells into the serum or tissue effusions (11, 14, 15).
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Entrez Gene IDs:
10232 (Human); 56047 (Mouse); 60333 (Rat)
CAK1 antigen; CAK1; megakaryocyte potentiating factor; mesothelin; MPF; MPFSMRP; MSLN; Pre-pro-megakaryocyte-potentiating factor; SMR; soluble MPF mesothelin related protein
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