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.

BioLegend’s Go-ChIP-Grade™ kit offers all the major components required for completing your ChIP assay from cell collection to immunoprecipitation and DNA purification. The kits feature spin columns based on solid-state technology from Chromatrap® and are suitable for ChIP assays from as little as 1 µg to up to 50 µg of protein. Our kit has been QC tested and validated with ChIP-qPCR.

Chromatrap’s Technology: This patented format is unique and has been granted in the UK (Patent No. GB2482209), the US (Patent No. 9523681), China (Patent No. ZL 2011 8 0067254.X) Japan (Patent No. JP 6088434) and Australia (Patent No. AU 2011340263).

Advantages of the kit:

  • The kit is compatible for use with ChIP-qPCR, ChIP-on-chip, and ChIP-seq assays
  • Suitable for ChIP assays using as little as 1 µg and to up to 50 µg of chromatin
  • ChIP assay can be completed in 1-2 days
  • Column contains inert polymer disc, reducing non-specific binding
  • Provides reduced immunoprecipitation (IP) incubation times
  • Elution chemistry is optimized for high quality and quantity of immunoprecipitated (IP'ed) DNA

 

Go-ChIP-Grade™ Protein G Enzymatic Kit

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Go-ChIP-Grade™ Protein G Enzymatic Kit provides a sensitive, reliable, and efficient method for enzymatic Chromatin IP. The kit is complete with all reagents and buffers necessary for ChIP assay and it revolutionizes the use of spin columns for ChIP assay. Each column contains a disc of an inert, porous polymer to which protein G has been covalently attached. During the assay, the target chromatin/antibody complex is retained by the disc. Flushing with three wash buffers and an elution step are all that is required to obtain the DNA fragments of interest. 

Go-ChIP-Grade™ Protein G Enzymatic Kit is preferable to other methods because:

  • Go-ChIP-Grade™ Protein G Enzymatic Kit columns eliminate the needs for beads.
  • Go-ChIP-Grade™ Protein G Enzymatic Kit columns contain a proprietary porous polymer disc to which protein G has been covalently anchored. Its pores have inner surfaces specifically designed to maximize chromatin capture efficiency.
  • The disc base material is chemically inert, reducing incidences of non-specific binding.
  • All chromatin capture takes place inside the disc, therefore there is no dependence on risky pipette separation procedures.
  • The standard protocol does not require pre-blocking and pre-wash steps.
  • Flushing away unbound chromatin and other unwanted material is easily achieved by using washing steps that are fast and less prone to error.
     

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