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Product Usage Information
For optimal ChIP and ChIP-seq results, use 2.5 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. The CUT&RUN dilution was determined using the CUT&RUN Assay Kit #86652. The CUT&Tag dilution was determined using 100,000 cells in a total reaction volume of 100 μl.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunofluorescence (Immunocytochemistry) | 1:100 - 1:400 |
| Chromatin IP | 1:200 |
| Chromatin IP-seq | 1:200 |
| CUT&RUN | 1:200 |
| CUT&Tag | 1:200 |
Specificity/Sensitivity
Species Reactivity:
Human, Mouse, Rat
参考图片
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or tunicamycin-treated (2 μg/ml, 8 hr; right), using ATF-4 (D4B8) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
共聚焦免疫荧光分析HeLa 细胞，未处理(左图)或衣霉素处理 (2 μg/ml, 8 hr; 右图)，所用抗体为 ATF-4 (D4B8) Rabbit mAb 兔单抗(绿色)。肌动蛋白微丝用 DY-554 phalloidin (红色)标记。
Western blot analysis of extracts from 293 and HeLa cells, untreated (-) or tunicamycin-treated (2 μg/ml, 8 hr; +), using ATF-4 (D4B8) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot方法检测293和HeLa细胞提取物：未经处理 (-) 和衣霉素处理(2 μg/ml, 8 hr; +)，使用的抗体是ATF-4 (D4B8) Rabbit mAb兔单抗 (上图) 或 β-Actin (D6A8) Rabbit mAb兔单抗 #8457 (下图)。
Immunoprecipitation of ATF-4 from extracts of 293 cells, treated with tunicamycin (2 μg/ml, 8 hr), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ATF-4 (D4B8) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ATF-4 (D4B8) Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as secondary antibodies.
免疫沉淀方法检测293细胞提取物ATF-4蛋白，使用衣霉素处理(2 μg/ml, 8 hr)，使用抗体为Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (泳道2)或ATF-4 (D4B8) Rabbit mAb (泳道3)。泳道1为10%对照。使用ATF-4 (D4B8) Rabbit mAb 进行Western blot方法分析，二抗是Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 和Anti-mouse IgG, HRP-linked Antibody #7076。
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MEF wild-type cells treated with tunicamycin (2ug/ml) overnight, and 5 µl of ATF-4 (D4B8) Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse ATF-3 Intron 1 Primers #13059, mouse CHOP promoter primers, 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.