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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. The CUT&RUN dilution was determined using 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 | 1:25 |
Immunohistochemistry (Paraffin) | 1:800 |
Immunofluorescence (Immunocytochemistry) | 1:400 |
Flow Cytometry (Fixed/Permeabilized) | 1:200 |
Chromatin IP | 1:50 |
Chromatin IP-seq | 1:50 |
CUT&RUN | 1:50 |
CUT&Tag | 1:50 |






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






参考图片
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with TSA #9950 (right), using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
使用Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb兔单抗 (绿色),共聚焦免疫荧光分析HeLa细胞,细胞分为untreated (左图)和TSA-treated (#9950;右图)。DY-554 phalloidin标记微丝蛋白(红色)。
Western blot analysis of lysates from HeLa and NIH/3T3 cells, untreated or TSA-treated (400 nM for 18 hours) using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb.
使用Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb兔单抗,免疫印迹(Western blot)分析HeLa和NIH/3T3细胞中Acetyl-Histone H3 (Lys9)的蛋白水平,细胞分为untreated或TSA-treated (400 nM for 18 hours)。
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Acetyl-Histone H3 (Lys9) (C5B11) 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 RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, 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,用4 x 106 HeLa细胞的交联染色质以及10 µl Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human RPL30 Exon 3 Primers #7014、SimpleChIP® Human GAPDH Exon 1 Primers #5516、SimpleChIP® Human MyoD1 Exon 1 Primers #4490和SimpleChIP® Human α Satellite Repeat Primers #4486,浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号,这相当于一。
Flow cytometric analysis of untreated HeLa cells using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 versus propidium iodide (DNA content). Note positive staining in cycling cells (box).
与propidium iodide (DNA含量)比较,使用Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649兔单抗,流式细胞仪分析未处理的HeLa细胞。显示在细胞周期中呈阳性染色 (方框)。
Immunohistochemical analysis of paraffin-embedded human gastic carcinoma using Acetyl-Histone H3 (K9) Rabbit mAb in the presence of non-acetyl-peptide (left) or K9 acetyl-peptide (right).
使用Acetyl-Histone H3(K9) Rabbit mAb兔单抗,免疫组化分析人胃癌组织石蜡切片,左图是非乙酰化的肽段,右图是K9乙酰化的肽段。