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Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ile86 of human NRF2 protein.
Product Usage Information
For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:10 - 1:50 |
| Chromatin IP | 1:50 |
Specificity/Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
参考图片
Chromatin immunoprecipitations were performed with cross-linked chromatin from MEF NRF2 wild-type cells (left) and NRF2 knockout cells (right), both treated with diethyl maleate (50 μM, 3 hr), and NRF2 (E5F1A) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse NQO1 Promoter Primers #12635 and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Western blot analysis of extracts from various cell lines, untreated (-) or treated with arsenite (50 μM, 8 hr; +), using NRF2 (E5F1A) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Simple Western™ analysis of lysates (1.0 mg/mL) from a172 cells treated with Mg132 (10uM, O/N) using NRF2 (E5F1A) Rabbit mAb #20733. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.