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Protein-DNA interactions play a critical role for cellular functions such as signal transduction, gene transcription and epigenetic silencing. In mammalian cells, interactions between the DNA-binding proteins and cognate promoter sequences are primary determinants in establishing spatial and temporal expression patterns of gene that affect homeostasis, development, and adaptation.
Abcam's ChIP Kit - One Step (ab117138) enables the user to enrich a chromatin fraction containing specific DNA sequences from isolated chromatin to be able to investigate the interaction between proteins and DNA. For best result, we recommend the use of Chromatin Extraction Kit (ab117152) for sample preparation.
|Components||Identifier||48 tests||96 tests|
|10X Wash Buffer||1 x 10ml||1 x 20ml|
|8-Well Assay Strips (with Frame)||6 units||12 units|
|8-Well Strip Caps||6 units||12 units|
|96-Well PCR Plate||1 unit||1 unit|
|Adhesive Covering Film||1 unit||1 unit|
|Anti-RNA Polymerase II (1 mg/ml)||1 x 8µl||1 x 16µl|
|ChIP Buffer||1 x 6ml||1 x 12ml|
|DNA Release Buffer||ECP3||1 x 14ml||1 x 28ml|
|GAPDH Primer - Forward (20 µM)||1 x 8µl||1 x 16µl|
|GAPDH Primer - Reverse (20 µM)||1 x 8µl||1 x 16µl|
|Non-Immune IgG (1 mg/ml)||1 x 10µl||1 x 20µl|
|Proteinase K (10 mg/mL)||1 x 55µl||1 x 110µl|
|Sonication Frame||1 unit||1 unit|
Our Abpromise guarantee covers the use of ab117138 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration.|
Chromatin was prepared from HeLa cells following Abcam's X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. ChIP was performed with ab8580 (Histone H3 tri-methyl K4) using ab117138. No antibody was added to the blank (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primes and probes are located in the first Kb of the transcribed region.
The analysis of enrichment of RNA polymerase II in GAPDH and MLH1 promoters by Abcam's ChIP Kit - One Step, with chromatin extract prepared from formaldehyde fixed colon cancer cells. Captured DNA was used for analyzing levels of RNA polymerase II enriched in the GAPDH and MLH1 promoters.
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