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Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly700 of human Stat3 protein.
Product Usage Information
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:100 |
| Immunofluorescence (Immunocytochemistry) | 1:1000 |
| Flow Cytometry (Fixed/Permeabilized) | 1:400 - 1:1600 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 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.
For a carrier free (BSA and azide free) version of this product see product #91276.
参考图片
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells starved overnight and treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/ml) for 30 minutes, and either Stat3 (D3Z2G) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IRF-1 promoter primers, SimpleChIP® Human c-Fos Promoter Primers #4663, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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 using Stat3 (D3Z2G) Rabbit mAb (upper), or β-Actin (D6A8) Rabbit mAb #8457 (lower). PC-3 cells are negative for Stat3.
Immunoprecipitation of Stat3 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Stat3 (D3Z2G) Rabbit mAb (lane 3). Lane 1 is 10% input.
Confocal immunofluorescent analysis of serum-starved HeLa cells, untreated (upper left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/ml, 30 min; upper right), and serum-starved PC-3 cells, untreated (lower right) or treated with Human Interferon-α1 (hIFN-α1) #8927 (100 ng/ml, 30 min; lower right), using Stat3 (D3Z2G) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Flow cytometric analysis of PC-3 cells (blue) and HeLa cells (green) using Stat3 (D3Z2G) Rabbit mAb or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells starved overnight and treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/ml) for 30 minutes and Stat3 (D3Z2G) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Fos, a known target gene of Stat3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells starved overnight and treated with Human Interleukin-6 (hIL-6) #8904 (100 ng/ml) for 30 minutes and Stat3 (D3Z2G) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 14 (upper), including Fos (lower), a known target gene of Stat3 (see additional figure containing ChIP-qPCR data).