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Anti-H4K8ac Polyclonal Antibody
Anti-H4K8ac Polyclonal Antibody
Artikelnummer
BPS25285
Verpackungseinheit
50 µg
Hersteller
BPS Bioscience
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Applications: ChIP (1 µg/IP)
ELISA (1:100)
DB (1:20,000)
WB (1:200)
IF (1:500)
Assay Conditions: ChIP results obtained with the antibody directed against H4K8ac
ChIP assays were performed using human HeLa cells, the antibody directed against H4K8ac (cat. No. 25285) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 2, 5, and 10 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the c-fos promoter and for the Sat2 satellite repeat region. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K8ac (cat. No. 25285), crude serum and flow through in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:16,700.Cross reactivity test using the antibody directed against H4K8ac
A Dot Blot analysis was performed to test the cross reactivity of the antibody against H4K8ac (cat. No. 25285) with peptides containing other histone modifications and the unmodified H4K8. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.Western blot analysis using the antibody directed against H4K8ac
Histone extracts of HeLa cells (15 µg) were analysed by Western blot using the antibody directed against H4K8ac (cat. No. 25285) diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.Immunofluorescence using the antibody directed against H4K8ac
Mouse NIH3T3 cells were stained with the antibody against H4K8ac (cat. No. 25285) and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H4K8ac 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.
Background: Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.
Concentration: 50 µg/41 µl
Description: Polyclonal antibody raised in rabbit against the region of histone H4 containing the acethylated lysine 20 (H4K8ac), using a KLH-conjugated synthetic peptide.
Format: Aqueous buffer solution
Formulation: PBS containing 0.05% azide and 0.05% ProClin 300
Immunogen: synthetic peptide
Purification: Affinity purified
Storage Stability: Store at -80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at -20°C for at least one month.
Uniprot: P62805
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
Applications: ChIP (1 µg/IP)
ELISA (1:100)
DB (1:20,000)
WB (1:200)
IF (1:500)
Assay Conditions: ChIP results obtained with the antibody directed against H4K8ac
ChIP assays were performed using human HeLa cells, the antibody directed against H4K8ac (cat. No. 25285) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 2, 5, and 10 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the c-fos promoter and for the Sat2 satellite repeat region. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K8ac (cat. No. 25285), crude serum and flow through in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:16,700.Cross reactivity test using the antibody directed against H4K8ac
A Dot Blot analysis was performed to test the cross reactivity of the antibody against H4K8ac (cat. No. 25285) with peptides containing other histone modifications and the unmodified H4K8. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.Western blot analysis using the antibody directed against H4K8ac
Histone extracts of HeLa cells (15 µg) were analysed by Western blot using the antibody directed against H4K8ac (cat. No. 25285) diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.Immunofluorescence using the antibody directed against H4K8ac
Mouse NIH3T3 cells were stained with the antibody against H4K8ac (cat. No. 25285) and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H4K8ac 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.
Background: Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.
Concentration: 50 µg/41 µl
Description: Polyclonal antibody raised in rabbit against the region of histone H4 containing the acethylated lysine 20 (H4K8ac), using a KLH-conjugated synthetic peptide.
Format: Aqueous buffer solution
Formulation: PBS containing 0.05% azide and 0.05% ProClin 300
Immunogen: synthetic peptide
Purification: Affinity purified
Storage Stability: Store at -80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at -20°C for at least one month.
Uniprot: P62805
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
| Artikelnummer | BPS25285 |
|---|---|
| Hersteller | BPS Bioscience |
| Hersteller Artikelnummer | 25285 |
| Green Labware | Nein |
| Verpackungseinheit | 50 µg |
| Mengeneinheit | STK |
| Reaktivität | Human, Mouse (Murine) |
| Klonalität | Polyclonal |
| Methode | Immunoprecipitation, Western Blotting, ELISA, Dot Blot, Chromatin Immunoprecipitation (ChIP) |
| Produktinformation (PDF) |
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| MSDS (PDF) |
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Immer für Sie da
Gernot Ensinger, M.Sc.
Laurent Haze
