Mesothelin Antibody (618923) [Unconjugated]

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.

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.

Recombinant Human Mesothelin protein was serially diluted 2-fold and captured by Mouse Anti-Human Mesothelin Monoclonal Antibody (Catalog # MAB32653) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human Mesothelin Monoclonal Antibody (Catalog # MAB32654) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

• Reactivity: Hu
• Applications: WB, Simple Western, ELISA
• Clone: 618923
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Mesothelin Antibody (618923) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Mesothelin
  Glu296-Asp580
  Accession # AAH09272
• Specificity: Detects human Mesothelin in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human MPF is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: MSLN
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • ELISA
  • Simple Western 20 ug/mL
  • Western Blot 2 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Mesothelin Antibody (618923) [Unconjugated]

• CAK1 antigen
• CAK1
• megakaryocyte potentiating factor
• Mesothelin
• MPF
• MPFSMRP
• MSLN
• Pre-pro-megakaryocyte-potentiating factor
• SMR
• soluble MPF mesothelin related protein

Background

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).

1. Hassan, R. et al. (2004) Clin. Cancer Res. 10:3937.
2. Kojima, T. et al. (1995) J. Biol. Chem. 270:21984.
3. Chang, K. and I. Pastan (1996) Proc. Natl. Acad. Sci. 93:136.
4. Onda, M. et al. (2006) Clin. Cancer Res. 12:4225.
5. Yamaguchi, N. et al. (1994) J. Biol. Chem. 269:805.
6. Muminova, Z.E. et al. (2004) BMC Cancer 4:19.
7. Hou, L.-Q. et al. (2008) Develop. Growth Differ. 50:531.
8. Ordonez, N.G. (2003) Mod. Pathol. 16:192.
9. Argani, P. et al. (2001) Clin. Cancer Res. 7:3862.
10. Li, M. et al. (2008) Mol. Cancer Ther. 7:286.
11. Scholler, N. et al. (1999) Proc. Natl. Acad. Sci. 96:11531.
12. Uehara, N. et al. (2008) Mol. Cancer Res. 6:186.
13. Rump, A. et al. (2004) J. Biol. Chem. 279:9190.
14. Ho, M. and M.O. Lively (2006) Cancer Epidemiol. Biomarkers Prev. 15:1751.
15. Robinson, B.W.S. et al. (2003) Lancet 362:1612.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Mesothelin Antibody (MAB32653)(2)

Publications using MAB32653

• Diane Manry, Kristian Bolanos, Breanna DiAndreth, Jee-Young Mock, Alexander Kamb Robust In Vitro Pharmacology of Tmod, a Synthetic Dual-Signal Integrator for Cancer Cell Therapy Frontiers in Immunology 2022-03-10 [PMID: 35359952]
• Elie Dheilly, Valéry Moine, Lucile Broyer, Susana Salgado-Pires, Zoë Johnson, Anne Papaioannou et al. Selective Blockade of the Ubiquitous Checkpoint Receptor CD47 Is Enabled by Dual-Targeting Bispecific Antibodies Molecular Therapy 2017-02-01 [PMID: 28153099] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: MSLN
• Uniprot:
  • AAH09272()