SMAD1 (phospho-S206) antibody

• Catalog Number: orb345659
• Category: Antibodies
• Description: SMAD1 (phospho-S206) antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a phosphorylated synthetic peptide corresponding to the region of amino acids containing serine 206 of human SMAD1 protein.
• Concentration: 0.82 mg/ml
• Dilution range: ELISA: 1:5,000 - 1:25,000, WB: 1:500 - 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This affinity-purified antibody is directed against the phosphorylated form of human SMAD1 protein at the pS206 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human SMAD1 pS206 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human SMAD1 is minimal by ELISA and western blot. A BLAST analysis was used to suggest 100% cross reactivity with SMAD1 from human, Duckbill platypus, Drosophila, mouse, opossum, chimpanzee, bonobo, orangutan, gorilla, macaque, cattle, sheep, pig, rat, horse, salmon, chicken, and dog based on the sequence homology with the immunogen. Reactivity against homologues from other sources is not known.
• Conjugation: Unconjugated
• UniProt ID: Q15797
• NCBI: Q15797.1
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-SMAD1 pS206 antibody, SMAD-1, SMAD 1,
• Note: For research use only
• Application notes: This affinity purified antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 52 kDa in size corresponding to phosphorylated SMAD1 protein by western blotting in the appropriate stimulated tissue or cell lysate or extract. This antibody is specific for the phosphorylated pS206 of SMAD1. Stimulation of EGF, TGFbeta BMP2, and 0.5M NaCl are recommended for 1 hour.
• Expiration Date: 12 months from date of receipt.

Western blot using Biorbyt's Affinity Purified anti-SMAD1 pS206 antibody shows detection of endogenous phosphorylated SMAD1 in whole cell lysates from human hepatoma (HEPG2, lanes 1-4) derived cell lines treated with PBS, BMP2 (5 ng/mL), EGF (1 ng/mL), or NaCl for 1 h at 37°C before harvest. Each lane contains approximately 15 µg of lysate. Primary antibody was used at a 1:500 dilution in 1% BLOTTO (p/n orb348624) and reacted for 1 hour at room temperature. Primary antibody was pre-incubated before reacting with blot as follows: top row - with PBS, middle row - with the immunizing phosphorylated peptide and bottom row - with control or non-phosphorylated peptide. The membrane was washed and reacted with a 1:3000 dilution HRP-conjugated a-Rabbit IgG (p/n orb347654) for 1 hour at room temperature. Detection was by ECL.