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Polyclonal antibody raised in rabbit against histone variant H2A.Z, using a KLH-conjugated synthetic peptide containing a sequence from the C-terminal part of the protein.
Lot
001D
Concentration
1 µg/µl
Species reactivity
Human: positive. Other species: not tested
Type
Monoclonal ChIP-grade ChIP-seq-grade
Purity
Affinity purified polyclonal antibody
Host
Rabbit
Storage Conditions
Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage Buffer
PBS containing 50% glycerol, 1% BSA and 0.09% azide
Precautions
This product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications
Suggested dilution
References
ChIP/ChIP-seq *
0.5 µg per IP
Fig 1, 2
Western Blotting
1:200
Fig 3
Immunofluorescence
1:100
Fig 4
*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5 - 5 µg per IP.
Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H2A.Z ChIP was performed with the Diagenode antibody against H2A.Z (cat. No. C15210021) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2.5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the EIF2S3 and EIF4A2 promoters, used as positive controls, and for the MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
A.
B.
C.
D.
Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H2A.Z ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H2A.Z (cat. No. C15210021) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the H2A.Z signal along the complete sequence and 1.5 Mb region of the human X-chromosome (figure 2A and B), and in two genomic regions surrounding the EIF2S3 and EIF4A2 positive controls (figure 2C and D).
Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H2A.Z Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H2A.Z (cat. No. C15210021). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.
Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H2A.Z HeLa cells were stained with the Diagenode antibody against H2A.Z (cat. No. C15210021, red) diluted 1:100. Actin was stained with fluorescein phalladoin (green).
ChIP-seq
Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on ... 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
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
