This antibody specifically recognizes shc/p66 when it is phosphorylated at Serine 36. We recommend to Immunoprecipitate shc/p66 prior to detection with mab shc/p66-6E10.
Characteristics
Synonyms: SHC-transforming protein 1, SHC-transforming protein A, SHC-transforming protein 3,SHCA, Src homology 2 domain-containing-transforming protein C1
Purification
Subsequent Thiophilic Adsorption and Size Exclusion Chromatography.
Western Blot: 1 μg/mL for HRPO/ECL detection. Recommended blocking buffer: Casein/Tween 20 based blocking and blot incubationbuffer. Positive Control: shc/p66 precipitated from EGF-treated HepG2 cells (See included Protocolbelow). ELISA. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
Positive Control Cell Lysate: HepG2 shc/p66-IPFormulationshc/p66 immunoprecipitated with anti-shc from EGF stimulated HepG2 cells. Proteins wereeluted with SDS and lyophilized from PBS/SDS. ReconstitutionReconstitute by addition of 125 µl H2O. After complete solubilization add 125 µl 2xSDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Aliquote and store frozen. Avoid repeated freezing and thawing.
Restrictions
For Research Use only
Reconstitution
Restore with 1 mL H2O (15 min, RT).
Buffer
1 mL2 x PBS, 0.09 % Sodium Azide, PEG and Sucrose
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
-20 °C/-80 °C
Storage Comment
Store lyophilized (preferably in a desiccator) at -20 °C and reconstituted (aliquote andfreeze in liquid nitrogen) at -80 °C. Avoid repeated freezing and thawing. Thaw aliquots at 37 °C. Thawed aliquots may be stored at 2-8 °C up to 3 months. Shelf life: one year from despatch.
Mammalian cells can express three alternatively spliced isoforms of the shc adaptor protein: shc/p46, shc/p52 and shc/p66. shc/p66 contains a unique N-terminal protein domain. In addition to tyrosine phosphorylation of Tyr 239/240 and/or Tyr 317, shc/p66 is phosphorylated at serine 36, e.g. in response to EGF. Serine phosphorylation of shc/p66 impairs its ability to bind to the activated EGF receptor thus inhibiting EGF receptor downstream signalling pathways.Synonyms: SHC-transforming protein 1, SHC-transforming protein 3, SHC-transforming protein A, SHCA, Src homology 2 domain-containing-transforming protein C1