Shc, p66 (SHC1, Src Homology and Collagen, Src Homology 2, FLJ26504, p52SHC, p66SHC, SH2 Domain Protein C1, SHC, SHCA, SHC Transforming Protein 1)

The human Src homology and collagen (SHC) gene encodes for three proteins (p46, p52, p66). All three SHCs are known to be signaling adaptors that couple activated growth factor receptors to their signaling pathways. p52 and p46 are translated from the same mRNA using two different initiation codons while p66 is generated form a distinct mRNA. p52 and p46 are expressed in every cell type while p66 expression varies from cell type to cell type. Upon stimulation by tyrosine kinase receptors and cytoplasmic TKs, all three SHCs become phosphorylated. Once phosphorylated, p52, p46 and p66 recruit and form a stable complex with GRB2/SOS. p52 and p46 activate the Ras and MAP kinase pathway, while p66 is involved in a signal transduction pathway that regulates the cellular response to oxidative stress and life span.\n\nApplications:\nSuitable for use in Western Blot and Immunoprecipitation. Other applications have not been tested.\n\nRecommended Dilutions:\nWestern Blot: 0.5-2ug/ml detects Shc isoforms in RIPA lysates from human A431 carcinoma cells, and previous lots detected Shc in mouse 3T3 RIPA cell lysate. A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with S1010-90. Proteins were visualized using an IgG\n(HRP) goat anti-rabbit secondary antibody and a chemiluminescence detection system.\nImmunoprecipitation: 4ug immunoprecipitates SHC isoforms from a human A431 RIPA cell lysate.\nOptimal dilutions to be determined by the researcher.\n\nPositive Control Included:\nA0001-50: Non-stimulated A431 cell lysate. Add 2.5ul of 2 mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Use 20ug of reduced lysate per lane for minigels.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

S1010-90

Size

250ug

Applications

IP, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, before the addition of glycerol to 30%.

P Type

Pab

Purity

Purified by Protein A affinity chromatography.

Isotype

IgG

References

1. He, H.J., et al., J. Biol. Chem. 278: 27,096-27,104 (2003). 2. Crouch, M.F., et al., J. Cell. Biol. 152: 263-273 (2001). 3. Gaudry, C.A., et al., J. Biol. Chem. 276: 24,871-24,880 (2001). 4. Mograbi, B., et al., J. Biol. Chem. 276: 45,307-45,319 (2001). 5. Gotoh, N., et al., Proc. Natl. Acad. Sci. USA 91: 167-171 (1994). 6. Pelicci, F., et al., Cell 70: 93-104 (1992).

Additional Info

Recognizes the human 46kD, 52kD and 66kD SHC proteins. Species Crossreactivity: mouse and rat.