Analysis Note

ControlBreast tumor tissuePurified active kinase is Catalogue Number 14-488

Application

Research Sub CategoryPI3K, Akt, & mTOR Signaling

Research CategorySignaling

Western blot 1:100 (55 kDa)

Immunocytochemistry

Immunohistochemistry: 1:20 to 1:50 for acetone-fixed, fresh frozen tissue. Not effective for paraffin sections.

Immunoprecipitation 1:10 to 1:20

Protocol for immunohistochemistry of frozen tissue sections on glass slides.

1. Fix sections for 5 minutes with cold acetone.

2. Air dry

3. Wash sections with PBS

4. Add blocking solution (horse serum) for 20 minutes

5. Add diluted PTEN antibody and incubate for 1 hour at room temperature.

6. Wash with PBS

7. Add biotinylated secondary antibody (horse anti-mouse, or equivalent).

8. Incubate for 30 minutes at room temperature.

9. Wash with PBS

10. Detect with ABC (Avidin-Peroxidase-Complex) or equivalent.

11. Wash with PBS

12. Develop with DAB substrate.

13. Hematoxylin dye may be used as a counterstain for visualizing nuclei.Breast tumor tissue gives good staining. Prostate tissue does not stain.Optimal working dilutions must be determined by end user.

Anti-PTEN Antibody, C-terminus, clone A2b1 is an antibody against PTEN for use in IC, IH, IP & WB.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Immunogen

Anti-PTEN was generated by immunization with a 10 amino acid peptide corresponding to amino acids 387-400 near the C-terminus of human PTEN.

Epitope: C-terminus

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-409

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein A purified

Format: Purified

Protein A Purified mouse immunoglobulin in 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative.

Specificity

Specifically recognizes PTEN*, a tumor suppressor protein located on human chromosome 10 (Steck, ′97; Li, ′97; Rhei, ′97) that is often mutated in various types of advanced cancers (Rhei, ′97). The PTEN protein exhibits protein phosphatase activity (Myers, ′97) and can suppress the growth of glioma cells (Furnari, ′97). The antibody recognizes human and mouse PTEN and is specific for the C-terminus. Other species not tested. Reactivity with human fibroblasts, breast epithelial and lung cancer cell lines has been confirmed by western blot, and antibody specificity has been confirmed by peptide blocking studies.

*PTEN is covered under U.S. patents 6,262,242, 6,482,795 and U.S. patent application 10/299,003.

Storage and Stability

Maintain for 1 year at 2–8°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Target description

55 kDa