Recombinant Human Mesothelin Fc Chimera Avi-tag Protein, CF

When Recombinant Human CA125/MUC16 (5609-MU) is coated at 0.2 μg/mL (100 μL/well), Biotinylated Recombinant Human Mesothelin C-terminal (aa 296‑580) Fc Chimera Avi‑tag (Catalog # AVI10679) binds with an ED50 of 2-12 ng/mL.

2 μg/lane of Biotinylated Recombinant Human Mesothelin His-tag Avi-tag (Catalog # AVI10679) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 70-78 kDa and 140-156 kDa,respectively.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Mesothelin Fc Chimera Avi-tag Protein, CF Summary

• Additional Information: Biotinylated, C-Terminal (aa 296-580)
• Details of Functionality: The biotin to protein ratio is greater than 0.7 as determined by the HABA assay. Measured by its binding ability in a functional ELISA. When Recombinant Human CA125/MUC16 (Catalog # 5609-MU) is coated at 0.2 μg/mL (100 μL/well), Biotinylated Recombinant Human Mesothelin C-terminal (aa 296‑580) Fc Chimera Avi‑tag binds with an ED₅₀ of 2-12 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Mesothelin protein
  

  Human Mesothelin (Glu296-Gly580) Accession # AAH09272.1	IEGRMD	Human IgG1 (Pro100-Lys330)	Avi-tag
  N-terminus			C-terminus

• Accession #: AAH09272.1
• N-terminal Sequence: Glu296
• Structure / Form: Disulfide-linked homodimer, biotinylated via Avi-tag
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 61 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 70-78 kDa, under reducing conditions

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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 1 mg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human Mesothelin Fc Chimera Avi-tag Protein, CF

• 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). This recombinant human Mesothelin lacks the 8 aa insertion, and within aa 296 - 580 it 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 up‑regulated 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). Our Avi-tag Biotinylated human Mesothelin features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

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.

Bioinformatics

• Uniprot:
  • AAH09272.1()