H3K36me3 polyclonal antibody - Classic

Name	H3K36me3 polyclonal antibody - Classic
Supplier	Diagenode
Catalog	C15310058
Prices	$295.00
Sizes	100 µl
Host	Rabbit
Clonality	Polyclonal
Applications	ChIPseq ChIP-PCR WB ELISA DB
Species Reactivities	Human
Purity/Format	Whole antiserum
Description	Rabbit Polyclonal
Supplier Page	Shop

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<strong> Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 μl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 μg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7

Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3
ChIP was performed with 5 μl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 μg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7

<strong> Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700.

Figure 3. Determination of the titer
To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700.

<strong> Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest.

Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3
A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest.