Recombinant Human Mesothelin Fc Chimera Avi-tag Protein, CF

Biotinylated, C-Terminal (aa 296-580)
Catalog # Availability Size / Price Qty
AVI10679-050
Biotinylated Recombinant Human Mesothelin His-tag Avi-tag Protein Binding Activity.
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Recombinant Human Mesothelin Fc Chimera Avi-tag Protein, CF Summary

Learn more about Avi-tag Biotinylated Proteins

Product Specifications

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
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
IEGRMDHuman IgG1
(Pro100-Lys330)
Avi-tag
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Glu296
Structure / Form
Disulfide-linked homodimer, biotinylated via Avi-tag
Predicted Molecular Mass
61 kDa
SDS-PAGE
70-78 kDa, under reducing conditions

Product Datasheets

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AVI10679

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI10679

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 1 mg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity View Larger

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.

SDS-PAGE View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Mesothelin

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.

References
  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.
Entrez Gene IDs
10232 (Human); 56047 (Mouse); 60333 (Rat); 102130460 (Cynomolgus Monkey)
Alternate Names
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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