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Anti-GDF15 Antibody 23B3D2.H5
Also for GDF15 (NM_004864)
|This Protein A purified antibody was prepared by repeated immunizations with a synthetic peptide corresponding to a region near the carboxy terminal end of human NAG-1 protein. A residue of cysteine was added to facilitate coupling to KLH.|
||ELISA: 1:2,000, WB: 1:1,000
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|Non-steroidal anti-inflammatory drug (NSAID) activated gene (NAG-1) is a member of the transforming growth factor-beta (TGF-beta) superfamily. NAG-1 is also known as Macrophage Inhibitory Cytokine-1 (MIC-1), Growth Differentiation Factor 15 (GDF15), Placental Bone Morphogenetic Protein (PLAB), or Prostate Derived Factor (PDF). NAG-1 is expressed in human placenta, prostate and colon. It possesses antitumorigenic and proapoptotic activities. NAG-1 expression is dramatically increased in inflammation, injury and malignancy. Increase of NAG-1 expression is a feature of many cancers including breast, colon, pancreas and prostate. In a number of studies, NAG-1 expression was increased by a number of NSAIDs. This increase in expression may correlate with the chemopreventive effect NSAIDs seem to have with certain cancers. NAG-1 expression is also induced by PPAR gamma ligands and by several dietary compounds such as conjugated linoleic acids (CLAs), naturally occurring fatty acids in ruminant food products, indoles, epicatechin gallate, and genistein. Induced expression of NAG-1 results in stimulation of apoptosis and inhibition of cell growth. Inhibition of NAG-1 induced expression by small interference RNA (siRNA) results in repression of induced apoptosis. NAG-1 expression is regulated by a numbers of transcription factors such as ERG-1 and Sp1. EGR-1 may be necessary for NSAID-induced NAG-1 expression. The study of expression of NAG-1 proteins, including variants, is important to define their potential role as serum biomarkers for cancer diagnosis, treatment monitoring, epidemiology study, and nutrition surveys.
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Western blot using anti-NAG-1 monoclonal antibody. The blot shows detection of recombinant NAG-1 protein present in Pichia pastoris whole cell lysates: lane 1 - yeast cell lysate expressing NAG-1 H variant with SUMO expression tag at 36 kDa; lane 2 - yeast cell lysate expressing NAG-1 D variant with SUMO expression tag at 36 kDa; lane 3 - yeast cell lysate expressing NAG-1 H variant; and lane 4 - yeast cell lysate expressing NAG-1 D variant. Recombinant NAG-1 proteins without SUMO correspond to monomer (15 kDa) and dimer (30 kDa) bands as indicated by the arrowheads. All lysates were run under reducing conditions. Primary antibody was used at a 1:1,000 dilution in TBS containg 1% BSA and 0.2% Tween, and reacted overnight at 4°C. For detection, a 1:40,000 dilution of peroxidase conjugated Gt-a-Mouse IgG secondary antibody (610-103-121) was used in Blocking Buffer for Fluorescent Western Blotting (MB-070) for 30 min at room temperature. Molecular weight estimation was made by comparison to prestained MW markers. Image was captured using the BioRad Versadoc™ 4000MP Imaging System.