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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) | 1:100 - 1:200 |
Flow Cytometry (Fixed/Permeabilized) | 1:100 |
Chromatin IP | 1:50 |
Chromatin IP-seq | 1:50 |






Specificity/Sensitivity
物种反应性:
人, 小鼠, 大鼠, 猴






参考图片
Western blot analysis of extracts from HT1080 (human), C2C12 (mouse) and B35 (rat) using Smad3 (C67H9) Rabbit mAb.
使用Smad3 (C67H9) Rabbit mAb兔单抗对HT1080 (人), C2C12 (小鼠)和B35 (大鼠)细胞提取物进行western blot分析。
Western blot analysis of extracts from HT1080 cells, treated with TGF-β1, TGFR inhibitor SB-431542 or BMP-2, using Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb #9520 (upper) or total Smad3 (C67H9) Rabbit mAb #9523 (lower).
使用Phospho-Smad3 (Ser423/425) (C25A9)Rabbit mAb兔单抗#9520 (上)或 total Smad3 (C67H9) Rabbit mAb兔单抗 #9523 (下)对经过 TGF-β1, TGFR 抑制因子 SB-431542 或BMP-2处理后的HT1080细胞提取物进行western blot分析。
Confocal immunofluorescent analysis of HT1080 cells, untreated (left) or TGFβ-treated (right), using Smad3 (C67H9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
使用Smad3 (C67H9) 兔单抗(绿色)对处理(左)或 TGFβ-处理(右)的HT1080细胞进行激光共聚焦荧光分析。使用DY-554 phalloidin(红色)标记肌动蛋白丝。
Flow cytometric analysis of HT-1080 cells using Smad3 (C67H9) Rabbit mAb #9523 (blue) compared to a nonspecific negative control antibody (red).
使用Smad3 (C67H9) Rabbit mAb兔单抗 #9523 对Ht-1080细胞进行流式细胞分析,使用非特异性抗体做为阴性对照(红色)。
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #3706 (7 ng/ml) for 1 h and either 10 μl of Smad3 (C67H9) 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® Human CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, 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.
使用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003对经过人TGF-β3 #3706 (7 ng/ml)处理的4 x 106 HaCaT细胞和10μl Smad3 (C67H9)Rabbit mAb兔单抗或2 μlNormal Rabbit IgG#2729进行染色质免疫共沉淀。富集到的DNA使用人CDKN1A内含子1引物#4669, SimpleChIP® 人ID1启动子引物#5139,人SMAD6启动子引物和SimpleChIP®人α Satellite Repeat引物#4486进行荧光实时PCR定量。各样品沉淀得到的DNA量比对于nput染色质量进行相对定量,Input的染色质量设定值为1。