Detection of Human Phospho-Smad2/3 (S465/S467) by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human TGF‑ beta 1 (Catalog # 240-B) for 30 minutes. PVDF membrane was probed with 2 μg/mL of Rabbit Anti-Human Phospho-Smad2/3 (S465/S467) Monoclonal Antibody (Catalog # MAB8935) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Smad2/3 (S465/S467) at approximately 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Smad2 (Sma and MAD-related Protein 2), also known as MAD2, MADR2 and MADH2, is a 58-62 kDa member of the Dwarfin/SMAD family of proteins. It is widely expressed, particularly in striated muscle, and exists constitutively in the cytoplasm. Smad2 is a downstream mediator of TGF-beta and activin signaling. In particular, Smad2 is a nonphosphorylated monomer in unstimulated cells. Upon ALK-4, -5, and -7 receptor activation, Smad2 is phosphorylated and forms either homotrimers or heterotrimers with Smad3 and Smad4. These heterotrimers enter the nucleus and initiate gene transcription. Human Smad2 is 467 amino acids (aa) in length. It contains one regulatory MH domain (aa 120-176) and a transactivation MH domain (aa 274-467). There are at least five utilized phosphorylation sites and one acetylation site at Lys19 that promotes transcriptional activity. There are four potential isoform variants. One shows a deletion of aa 79-108, while another contains the same deletion coupled to another deletion of aa 219-243. A third shows a deletion of aa 344-358, and a fourth is missing aa 221-225. Over aa 20-108, human Smad2 shares 98% aa identity with mouse Smad2.
Mothers Against DPP Homolog 2/3
hMAD-2; hSMAD2; JV18; JV18-1SMAD, mothers against DPP homolog 2 (Drosophila); MAD homolog 2; Mad protein homolog; Mad, mothers against decapentaplegic homolog 2; MADH2MGC34440; MADR2MGC22139; Mad-related protein 2; mother against DPP homolog 2; mothers against decapentaplegic homolog 2; Mothers against DPP homolog 2; Sma- and Mad-related protein 2; SMAD 2; SMAD family member 2MAD, mothers against decapentaplegic homolog 2 (Drosophila); SMAD, mothers against DPP homolog 2; Smad2; Smad2/3
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