Human Mesothelin Antibody

Catalog # Availability Size / Price Qty
MAB32653
MAB32653-SP
Detection of Human Mesothelin by Western Blot.
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Human Mesothelin Antibody Summary

Species Reactivity
Human
Specificity
Detects human Mesothelin in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human MPF is observed.
Source
Monoclonal Mouse IgG1 Clone # 618923
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Mesothelin
Glu296-Asp580
Accession # AAH09272
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Simple Western
20 µg/mL
See below
ELISA

This antibody functions as an ELISA capture antibody when paired with Mouse Anti-Human Mesothelin Monoclonal Antibody (Catalog # MAB32654).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Mesothelin DuoSet ELISA Kit (Catalog # DY3265) for convenient development of a sandwich ELISA or the Human Mesothelin Quantikine ELISA Kit (Catalog # DMSLN0) for a complete optimized ELISA.

 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Mesothelin antibody by Western Blot. View Larger

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.

Simple Western Detection of Human Mesothelin antibody by Simple Western<sup>TM</sup>. View Larger

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.

Human Mesothelin ELISA Standard Curve. 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).

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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
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.

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). 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).

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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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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Human Mesothelin Antibody
By Anonymous on 12/07/2020
Application: MiAr Sample Tested: EDTA Plasma Species: Human

Antibody was printed on custom arrays and incubated with fluorescently labeled human EDTA plasma