PhosphoDetect Anti-Fos (pSer374) Mouse mAb (34E4)

Analysis Note

Positive ControlHepG2 cells treated with EGF

Application

ELISA (0.1 g/ml)Immunoblotting (0.5 g/ml, chemiluminescence)

General description

Mouse monoclonal antibody purified from serum-free cell culture supernatant by thiophilic adsorption and size exclusion chromatography. Supplied with a positive control lysate consisting of EGF-treated HepG2 cells. Recognizes the ~55 kDa c-fos protein phosphorylated at Ser³⁷⁴ by MAP kinase.

This PhosphoDetect™ Anti-Fos (pSer³⁷⁴) Mouse mAb (34E4) is validated for use in ELISA, Immunoblotting for the detection of Fos (pSer³⁷⁴).

The early gene product c-Fos is expressed following mitogenic stimulation and functions as a sensor for MAPK signal duration. When MAPK activation is transient, MAPK activity declines before accumulation of the c-Fos protein. When MAPK activation is sustained, c-Fos is phosphorylated by MAPK at Ser³⁷⁴. Phosphorylation stabilizes the Fos protein and primes c-Fos for additional phosphorylation at Thr³²⁵.

Recognizes the ~55 kDa c-Fos protein phosphorylated at Ser³⁷⁴. Does not detect the unphosphorylated form.

Immunogen

a synthetic phosphopeptide corresponding to amino acids surrounding the Ser³⁷⁴ phosphorylation site of human c-Fos

Human

Legal Information

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

Other Notes

Does not detect the unphosphorylated form of c-fos. For immunoblotting use 20 µl control lysate per lane (minigel) for chemiluminescent detection. Antibody should be titrated for optimal results in individual systems.

Chen, R-H., et al. 1993. Proc. Natl. Acad. Sci. USA90, 10952.

Packaging

1 set in Plastic ampoule

Physical form

100 µg antibody lyophilized from 2X PBS, PEG, sucrose and 200 µl lyophilized control lysate from EGF-treated HEPG2 cells.

Reconstitution

Reconstitute antibody with 1 ml water (15 minutes, room temperature). Following reconstitution, aliquot and freeze in liquid nitrogen. Reconstituted antibody can be stored at -80°C for up to 1 year. Aliquots may be stored at 4°C for up to 3 months and should be thawed at 37°C. Reconstitute control lysate with 200 µl H₂O. After complete solubilization of the proteins add 200 µl SDS-PAGE sample buffer and incubate at 90°C for 5 min. Following reconsitution aliquot and freeze (-20°C). Avoid freeze/thaw cycles.

Warning

Toxicity: Irritant (B)