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H4K16ac Antibody
Catalog Number
Format
Price
Polyclonal antibody raised in rabbit against Histone H4 (ac Lys16), using a KLH-conjugated synthetic peptide.
| Lot | 001 |
|---|---|
| Concentration | 1 mg/ml |
| Species reactivity | Human, mouse, C. elegans, rat, chicken, Xenopus, Drosophila, plant: positive. Other species: not tested. |
| Type | Polyclonal, CUT&Tag-grade |
| Purity | Affinity purified |
| Host | Rabbit |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride; contains 0.01% azide as preservative |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| CUT&Tag | 1 µg | Figure 1 |
| Dot Blotting | 1:1,000 | Figure 2 |
| Western Blotting | 1:500 | Figure 3 |
| Immunofluorescence | 1:100 | Figure 4 |
- Validation Data
Figure 1. CUT&Tag results obtained with the antibody directed against H4K16ac
CUT&Tag was performed on 50,000 K562 cells using 1 µg of the antibody against H4K16ac (cat. No. C15410300) and the Universal CUT&Tag kit (cat. No. C01070024). The libraries were subsequently analysed on an Illumina NovaSeqX sequencer (2x150 bp paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg38) using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 2 Mb region of chromosome 5 (figure 1A and B) and in 2 genomic regions on chromosome 7 and 12 (figure 1C and D, respectively).
Figure 2. Cross reactivity test using the antibody directed against H4K16ac
A Dot Blot analysis was performed to test the cross reactivity of the antibody against H4K16ac (cat. No. C15410300) with peptides containing the acetylated or unmodified H4K16 sequence. One hundred to 1 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 2 shows a high specificity of the antibody for the acetylated peptide.
Figure 3. Western blot analysis using the antibody directed against H4K16ac
Western blot was performed on 30 µg histone extracts from HeLa (lane 1) and NIH-3T3 cells (lane 2) using the antibody against H4K16ac (cat. No. C15410300). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the right.
Figure 4. Immunofluorescence using the antibody directed against H4K16ac
HeLa cells were fixed with 0.5% formaldehyde and blocked with PBS/ TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H4K16ac antibody (upper left) diluted 1:100 in blocking solution followed by an FITC labelled anti-rabbit secondary antibody. The upper right panel shows staining of the nuclei with DAPI, whereas the lower left panel shows staining of alpha-tubulin with Dylight 594. A merge of the three stains is shown in the bottom right panel. - Publications
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