AM26533AF-N Mesothelin antibody

Swiss Prot Num:

Q61468

Immunogen:

Murine hemangioblast-like cell line LO

GeneID:

56047

Mesothelin (MSLN) is a GPI-linked cell surface glycoprotein expressed in the mesothelial lining of the body cavities and in mullerian duct epithelium related cancer cells (ovarian, pancreatic cancer, mesothelioma). Both human and mouse mesothelin bind to ovarian cancer antigen CA125/MUC16 with high affinity, mediating cell attachment in vitro , suggesting that mesothelin may facilitate ovarian cancer metastasis.

Flow cytometric analysis for floating cells
We usually use Fisher tubes or equivalents as reaction tubes for all steps described below.
1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN3].
2 ) Resuspend the cells with washing buffer (5x10e6 cells/mL).
3) Add 50 µL of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute at room temperature (20~25°C). Remove supernatant by careful aspiration.
4) Add 10 µL of normal goat serum containing 1 mg/mL normal human IgG and 0.1% NaN3 to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature.
5) Add 30 µL of the primary antibody at the concentration as suggested in the APPLICATIONS diluted in the washing buffer. Mix well and incubate for 30 minutes at room temperature.
6) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
7) Add 30 µL of 1:100 FITC conjugated anti-rat IgG diluted with the washing buffer. Mix well and incubate for 15 minut es at room temperature.
8) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
9) Resuspend the cells with 500 µ L of the washing buffer and analyze by a flow cytometer.
(Positive control for Flow cytometry; LO)

1. Gubbels, J. A., et al., Mol. Cancer 5 , 50-64 (2006).
2. Ho, M., et al., Clin. Cancer Res. 11 , 3814-3820(2005).
3. Rump, A., et al., J. Biol. Chem. 279 , 9190-9198(2004).
4. Nakayama, K., et al., J. Biol. Chem . 274 , 24766-24772 (1999).
5. Kojima, T., et al., J. Biol. Chem. 270 , 21984-21990(1995).

Purification:

Protein G Agarose Chromatography

Buffer System:

PBS containing 50% Glycerol, pH 7.2. No preservative is contained.

State:

Liquid purified Ig fraction
Azide Free

Species reactivity (tested):

Human (Jurkat), Mouse.

Specificity:

This antibody reacts with Mesothelin.