Description: Rabbit polyclonal antibody to ATF4 (Phospho-S245) Immunogen: KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding S245 of human ATF4 protein. The exact sequence is proprietary. Purification: The antibody was purified by immunogen affinity chromatography. Clonality: Polyclonal Form: Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide. Dilution: WB (1/500 - 1/1000), IH (1/50 - 1/100), IF/IC (1/50 - 1/200) Gene Symbol: ATF4 Alternative Names: CREB2; TXREB; Cyclic AMP-dependent transcription factor ATF-4; cAMP-dependent transcription factor ATF-4; Activating transcription factor 4; Cyclic AMP-responsive element-binding protein 2; CREB-2; cAMP-responsive element-binding protein 2; DNA-binding protein TAXREB67; Tax-responsive enhancer element-binding protein 67; TaxREB67Entrez Gene (Human): 468Entrez Gene (Mouse) : 11911Entrez Gene (Rat) : 79255SwissProt (Human): P18848SwissProt (Mouse) : Q06507SwissProt (Rat) : Q9ES19Storage/Stability : Shipped at 4°C. Upon delivery aliquot and store at -20°C for one year. Avoid freeze/thaw cycles.
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Western blot analysis of ATF4 (Phospho-S245) expression in SGC7901 (A), 3T3L1 (B), C6 (C), EC9706 (D), A375 (E) whole cell lysates.
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Immunohistochemical analysis of ATF4 (Phospho-S245) staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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Immunofluorescent analysis of ATF4 (Phospho-S245) staining in A549 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a AF488-conjugated secondary antibody (green) in PBS at room temperature in the dark. Phalloidin - AF594 was used to stain Actin filaments (red). DAPI was used to stain the cell nuclei (blue).
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Direct ELISA antibody dose-response curve using Anti-ATF4 (Phospho-S245) Antibody. Antigen (Phosphopeptide and non-phosphopeptide) concentration is 5 ug/ml. Goat Anti-Rabbit IgG (H&L) - HRP was used as the secondary antibody, and signal was developed by TMB substrate.