Go-ChIP-Grade™ antibodies are validated for use in ChIP experiments and compared to a competitor ChIP-validated antibody to ensure equal or better performance. Each lot is quality tested in ChIP-qPCR experiments. Additionally, many of our clones are validated for use in other applications including Western Blot, Immunofluorescence, and Immunohistochemistry. All our products are supported by our 100% satisfaction guarantee and our highly capable technical team can assist if you have any questions.

 

Our purified Go-ChIP-Grade™ antibodies are provided at concentrations between 0.5 mg/ml- 1.0 mg/ml, and our ascites antibodies are estimated to be between 1-3 mg/ml.

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Chromatin Immunoprecipitation (ChIP) was performed using commercial Protein-G coated 96 well high-throughput ChIP assay kit by loading 3µg of cross-linked chromatin samples from HeLa cells treated with Nocodazole with either A) 1:50 dilution of Go-ChIP-Grade™ Purified anti-STAT1 Phospho (Ser727) Antibody (Clone A15158B), or B) equal amount of Purified Mouse IgG1, κ Isotype Control Antibody, or C) competitor’s ChIP-grade Purified anti-STAT1 Phospho (Ser727) Antibody and D) equal amount of matched Isotype Control Antibody as recommended by the manufacturer. The enriched DNA was purified and quantified by real-time qPCR using primers targeting human IRF1 gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.

 

Total lysates (15µg protein) from untreated HeLa cells (lane 1) and HeLa cells treated with nocodazole (lane 2) were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 1:500 diluted (1µg/mL) Purified anti-STAT1 Phospho (Ser727) Antibody, clone A15158B (upper) or 1:3000 diluted Purified anti-β-actin Antibody, clone Poly6221 (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-STAT1 Phospho (Ser727) Antibody, and a donkey anti-rabbit IgG Antibody conjugated to HRP for anti-β-actin Antibody.

 

Human peripheral blood lymphocytes were stimulated with (filled histogram) or without (open histogram) Cell Activation Cocktail (without Brefeldin A) for 15 minutes, fixed with Fixation Buffer, permeabilized with True-Phos™ Perm Buffer, then intracellularly stained with purified anti-STAT1 Phospho (Ser727) antibody (clone A15158B), followed by anti- mouse IgG PE.

 

HeLa cells were stimulated with (filled histogram) or without (open histogram) nocodozole for 24 hours, fixed with Fixation Buffer, permeabilized with True-Phos™ Perm Buffer, then intracellularly stained with purified anti-STAT1 Phospho (Ser727) antibody (clone A15158B), followed by anti- mouse IgG PE.

 

Two aliquots of HeLa cells in suspension, treated with (left) 200 ng/mL nocodazole for 24 hours or left un-treated (right) were adhered on poly-lysine pre-coated slides. Then they were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for three minutes, and blocked with 5% FBS for 60 minutes. The cells were intracellularly stained with 2 µg/ml of purified anti-STAT1 Phospho (Ser727) antibody (clone A15158B) overnight at 4°C followed by Alexa Fluor®594 (red) conjugated goat anti-mouse IgG for one hour at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.