JARID1C (UniProtKB/Swiss-Prot entry P41229) is a histone demethylase that specifically demethylates tri- and dimethylated Lys-4 of histone H3. It has no activity towards monomethylated H3K4, H3K9, H3K27, H3K36, H3K79 or H4K20. It is involved in transcriptional regulation and chromatin remodeling. JARID1C participates in transcriptional repression of neuronal genes by recruiting histone deacetylases and REST at neuron-restrictive silencer elements. Mutations in JARID1C have been associated with X-linked mental retardation
JARID1C Antibody - ChIP-seq Grade (sample size)
Polyclonal antibody raised in rabbit against human JARID1C (Jumonji, AT Rich Interactive Domain 1C), using a synthetic peptide containing a sequence from the central part of the protein1.
| Lot | A301-034A2 |
|---|---|
| Concentration | 0.2 μg/μl |
| Species reactivity | Human: positive. Other species: not tested. |
| Type | Polyclonal |
| Purity | Affinity purified polyclonal antibody in TBS containing 0.1% BSA and 0.09% azide. |
| Host | Rabbit |
| Storage Conditions | Store at 4°C. |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq * | 5 μg/ChIP | Fig 1, 2 |
| Western Blotting | 1:100 | Fig 3 |
| IP | 6 μg per IP | Fig 4 |
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 μg per IP.
1Manufactured by Bethyl Laboratories, Inc., Texas, USA
- Validation data
Figure 1. ChIP results obtained with the Diagenode antibody directed against JARID1C
ChIP assays were performed using K562 cells, the Diagenode antibody against JARID1C (Cat. No. C15410338) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055), using sheared chromatin from 4 million cells. A titration consisting of 1, 2 and 5 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoter of the active EIF4A2 gene, used as positive control, and for the MYOD1 gene and Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against JARID1C
ChIP was performed on sheared chromatin from 4 million K562 cells using 5 μg of the Diagenode antibody against JARID1C (Cat. No. C15410338) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 600 Kb region of the human chromosome 1 (fig 2A and B), and in two genomic regions surrounding the ALDOA gene and the EIF4A2 positive control gene (fig 2C and D).
Figure 3. Western blot analysis using the Diagenode antibody directed against JARID1C
Whole cell extracts from HeLa cells were analysed by Western blot using the Diagenode antibody against JARID1C (Cat. No. C15410338) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
Figure 4. Immunoprecipitation using the Diagenode antibody directed against JARID1C
Immunoprecipitation was performed on whole cell extracts from HeLa cells using 6 μg of the Diagenode antibody against JARID1C (Cat. No. C15410338, lane 1). An equal amount of rabbit IgG was used as a negative control (lane 2). The immunoprecipitated JARID1C protein was detected by western blot with the JARID1C antibody diluted 1:200. - 出版物
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We strongly recommend using this: JARID1C Antibody - ChIP-seq Grade (sample size) (Hologic Diagenode Cat# C15410338-20 Lot# A301-034A2). Click here to copy to clipboard.
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