Chromatin Immunoprecipitation (ChIP) was performed using Go-ChIP-Grade™ Protein G Enzymatic Kit by loading 3 µg of cross-linked chromatin samples from HeLa cells with either A) 1:300 dilution of Go-ChIP-Grade™ Purified anti-RNA Polymerase II Antibody (clone 8WG16, Cat. No. 664911 & 664912), B) equal amount of Purified Mouse IgG2a Isotype Control Antibody (Cat. No. 400201), or C) competitor’s ChIP-grade Purified anti-RNA Polymerase II 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 GAPDH gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.
Chromatin Immunoprecipitation (ChIP) was performed using Go-ChIP-Grade™ Protein G Enzymatic Kit by loading 3 µg of cross-linked chromatin samples from HeLa cells with either A) 1:300 dilution of Go-ChIP-Grade™ Purified anti-RNA Polymerase II Antibody (clone 8WG16, Cat. No. 664911 & 664912), B) equal amount of Purified Mouse IgG2a Isotype Control Antibody (Cat. No. 400201), or C) competitor’s ChIP-grade Purified anti-RNA Polymerase II 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 GAPDH gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.
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Description
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.
Go-ChIP-Grade™ anti-RNA Polymerase II Antibody; clone 8WG16 (Positive control antibody)
50uL
4
Mouse IgG2a, k Isotype Control Antibody
50uL
4
Forward primer for GAPDH (human promoter region)
25uL
-20
Reverse primer for GAPDH (human promoter region)
25uL
-20
1.5 ml Collection Tubes
12
RT
DNA Purification Columns
6
RT
DNA Binding Buffer
3.75mL
RT
DNA Wash Buffer
3.75mL
RT
DNA Elution Buffer
500uL
RT
For more information on individual components, please refer to the product manual.
Chromatrap® 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).
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