HDAC1 Antibody
Alternative names: HD1, RPD3, RPD3L1, GON-10
Polyclonal antibody raised in rabbit against the C-terminal region of human HDAC1 (Histone deacetylase 1), using a KLH-conjugated synthetic peptide.
| Lot | A21-001P |
|---|---|
| Concentration | 1.73 μg/μl |
| Species reactivity | Human, mouse |
| Type | Polyclonal |
| Purity | Affinity purified |
| Host | Rabbit |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq * | 2 μg/ChIP | Fig 1, 2 |
| ELISA | 1:4,000 | Fig 3 |
| Western Blotting | 1:1,000 | Fig 4, 5 |
| Protein array | 1:100,000 | Fig 6 |
| Immunofluorescence | 1:500 | Fig 7 |
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.
Validation Data
Figure 1. ChIP results obtained with the Diagenode antibody directed against HDAC1
ChIP was performed with the Diagenode antibody against HDAC1 (Cat. No. C15410325) on sheared chromatin from 4,000,000 HeLa cells. An antibody titration consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and GAPDH promoters, used as positive controls, and for the MYOD1 gene and Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against HDAC1
ChIP was performed on sheared chromatin from 4,000,000 HeLa cells using 2 μg of the Diagenode antibody against HDAC1 (Cat. No. C15410325) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1 Mb region of the X-chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure 2C and D)..
Figure 3. Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against HDAC1 (Cat. No. C15410325), crude serum and flow through. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:75,000.
Figure 4. Western blot analysis using the Diagenode antibody directed against HDAC1
Whole cell extracts (25 μg, lane 1) and nuclear extracts (25 μg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against HDAC1 (Cat. No. C15410325) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.
Figure 5. Western blot analysis using the Diagenode antibody directed against HDAC1
Whole cell extracts (50 µg) from HeLa cells transfected with HDAC1 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against HDAC1 (Cat. No. C15410325) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.
Figure 6. Protein array analysis with the Diagenode antibody directed against HDAC1
The specificity of the Diagenode antibody against HDAC1 (Cat. No. C15410325) was demonstrated using the HuProt human protein microarray (CDI Laboratories), a protein array containing more than 19,000 human proteins. The antibody was used at a dilution of 1:100,000. Figure 6 shows the Z-score of the signal intensity (mean value of the duplicate spots on the array). The names of the proteins with 5 highest Z-scores are indicated at the bottom. This figure clearly shows the high specificity of the antibody for HDAC1.
Figure 7. Immunofluorescence using the Diagenode antibody directed against HDAC1
HeLa cells were stained with the Diagenode antibody against HDAC1 (Cat. No. C15410325) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the HDAC1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Target description
HDAC1 (UniProt/Swiss-Prot entry Q13547) catalyses the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Acetylation and deacetylation of these highly conserved lysine residues is important for the control of gene expression and HDAC activity is often associated with gene repression. Histone deacetylation is established by the formation of large multiprotein complexes. HDAC1 also interacts with the retinoblastoma tumor suppressor protein and is able to deacetylate p53. Therefore, it also plays an essential role in cell proliferation and differentiation and in apoptosi.
Applications
| ChIP-seq (ab) Read more |
| ELISA Enzyme-linked immunosorbent assay. Read more |
| WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more |
| Protein Analysis Proteins play a critical role in virtually all cell processes including metabolism, structure, growth, and repair. Understanding protein function, behavior, structure, and their importance are key aspects of understanding disease and therapeut... Read more |
| IF Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more |
| ChIP-qPCR (ab) Read more |
| siRNA Knockdown Epigenetic antibodies you can trust! Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level o... Read more |
Documents
| HDAC1 polyclonal antibody - Premium DATASHEET Polyclonal antibody raised in rabbit against the C-terminal region of human HDAC1 (Histone deacet... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
Safety sheets
Publications
How to properly cite our product/service in your workWe strongly recommend using this: HDAC1 Antibody (Hologic Diagenode Cat# C15410325 Lot# A21-001P). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
NUP98 and RAE1 sustain progenitor function through HDAC-dependentchromatin targeting to escape from nucleolar localization. |
Histone Deacetylases 1 and 2 target gene regulatory networks of nephronprogenitors to control nephrogenesis. |
Threonine phosphorylation of IκBζ mediates inhibition of selective proinflammatory target genes. |
Immunity drives regulation in cancer through NF-κB. |
Immunity drives TET1 regulation in cancer through NF-kB |
Krüppel-like transcription factor KLF10 suppresses TGFβ-induced epithelial-to-mesenchymal transition via a negative feedback mechanism |
HDAC1 negatively regulates Bdnf and Pvalb required for parvalbumin interneuron maturation in an experience-dependent manner |
Genome-wide hydroxymethylcytosine pattern changes in response to oxidative stress |
SNAIL1 combines competitive displacement of ASCL2 and epigenetic mechanisms to rapidly silence the EPHB3 tumor suppressor in colorectal cancer. |
Citrullination of DNMT3A by PADI4 regulates its stability and controls DNA methylation. |
Dimethyl fumarate regulates histone deacetylase expression in astrocytes. |
Phosphorylation of p65(RelA) on Ser547 by ATM Represses NF-κB-Dependent Transcription of Specific Genes after Genotoxic Stress |
The histone demethylase Kdm3a is essential to progression through differentiation. |
HDAC1 Regulates Fear Extinction in Mice. |
Enhancer of Zeste 2 (EZH2) is up-regulated in malignant gliomas and in glioma stem-like cells. |
The core binding factor CBF negatively regulates skeletal muscle terminal differentiation. |
Functional connection between deimination and deacetylation of histones. |