Members of the SMAD family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-beta signals from the cell surface into the nucleus. Three distinct classes of SMADs have been defined: the receptor-regulated SMADs (R-SMADs), which include Smad1, 2, 3, 5 and 8, the common-mediator SMAD (co-SMAD), SMAD4, and the antagonistic or inhibitory SMADs (I-SMADs), SMAD6 and 7. Once in the nucleus, SMADs can target a variety of DNA binding proteins to regulate transcriptional responses. Following stimulation by TGF-beta, SMAD2 and SMAD3 become phosphorylated at their carboxyl termini (Ser465 and 467 on SMAD2; Ser423 and 425 on SMAD3) by TGF-beta Receptor I. Phosphorylated SMAD 2/3 can complex with SMAD4, translocate to the nucleus and regulate gene expression.
Applications:
Suitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry, and Immunocytochemistry. Other applications not tested.
Recommended Dilution:
Western Blot: 1:5,000
Immunohistochemistry (paraffin): 1:100-250
Immunocytochemistry: 1:100-250
Immunoprecipitation: 1:50
Optimal dilutions to be determined by the researcher.
Positive Control:
Jurkat cell lysate
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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