Spike-in Chromatin
Catalog No: 53083
Lot No: 02317006
Format: 15 rxns
Quality Control: The Spike-in Chromatin is sonicated using Active Motif’s Epishear Probe sonicator. Chromatin is then
tested with Spike-in Antibody (Catalog No. 61686) in combination with human chromatin and a robust histone modification
antibody using Active Motif’s ChIP-IT High Sensitivity Kit (Catalog No. 53040). ChIP qPCR is performed using primers for
human and Drosophila genes. ChIP-seq results are mapped to the human and Drosophila reference genomes.
Contents
Spike-in Chromatin prepared from Schneider’s Drosophila Line 2 (S2) cells is provided for 15 rxns of robust histone
modification antibody targets. Chromatin is provided at a concentration of 10 ng/µl.
Robust Histone Modification Ab
Transcription Factor Ab
Sample Chromatin Spike-in Chromatin
25 µg
50 ng
25 µg
20 ng
Antibody of Interest
4 µg
4 µg
Spike-in Antibody
2 µg
2 µg
If optimizing the amount of chromatin to use per ChIP reaction, we recommend adjusting the amount of spike-in chromatin
used in order to maintain the same chromatin ratio (Sample:Spike-in) as suggested on the data sheet.
Background
The Spike-in Chromatin is intended for use with Active Motif’s Spike-in Antibody (Catalog No. 61686) to normalize
chromatin immunoprecipitation (ChIP) experiments. ChIP is a multi-step process in which variations caused by sample
loss during immunoprecipitation and library preparation, uneven sequencing read depth or user differences can lead to
results that are difficult to interpret. To overcome this challenge, Active Motif has developed a spike-in strategy to
normalize out technical variation and sample processing bias. Additionally, the normalization strategy can be used to
monitor the effects of experimental conditions, such as inhibitory compounds or mutants.
A standard ChIP reaction is set up using experimenal chromatin (e.g. human) and an antibody of interest. In addition,
Drosophila melanogaster chromatin is added, or “spiked-in”, to each reaction as a minor fraction of the total chromatin. An
antibody that recognizes the Drosophila-specific histone variant, H2Av, is also added to the reaction. The Spike-in
antibody provides a mechanism to reliably pull down a small fraction of Drosophila chromatin that is consistent across all
samples. Since variation introduced during the ChIP procedure will aslo occur with the spike-in chromatin, a normalization
factor can be created based on the Drosophila signal and applied to the sample genome. Unlike other normalization
methods that rely on the same protein-specific antibody for both sample and normalization control, Active Motif’s
normalization strategy with the separate Spike-in antibody enables normalization across different antibodies without bias.
Storage and Guarantee: Store at -80°C.
We recommend aliquoting the chromatin into single-use fractions and then storing them at -80°C. This eliminates
repeated freeze/thaw cycles. This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
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