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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Chromatin Immunoprecipitation [1]
- Other assay [2]
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- Product number
- PA5-16182 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- H4K8ac Polyclonal Antibody, ChIP-Verified
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on acid extracts of HeLa (Lane 1), HeLa treated with sodium butyrate (5mM Sodium butyrate for 24 hr) (Lane 2), HCT 116 (Lane 3) and HCT 116 treated with sodium butyrate (5mM Sodium butyrate for 24 hr) (Lane 4). The blot was probed with Anti-Acetyl Histone H4 (Lys8) (Product # PA5-16182, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 14 kDa band corresponding to Acetyl-Histone H4 (Lys8) was observed and was enhanced upon treatment across the cell lines tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Acetyl-Histone H4 (Lys8) was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Acetyl-Histone H4 (Lys8) Rabbit Polyclonal Antibody (Product # PA5-16182) at 1:250 dilution in 0.1% BSA, incubated overnight at 4 degree Celsius and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Enrichment of endogenous Acetyl-Histone H4 (Lys8) protein at specific gene loci using Anti-Acetyl-Histone H4 (Lys8) Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-Acetyl-Histone H4 (Lys8) Rabbit Polyclonal Antibody (Product # PA5-16182, 3 µg) on sheared chromatin from 2 million Sodium butyrate-treated HeLa cells using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over the ACTB gene (active) and MYOD and SAT2 satellite repeats (inactive). A schematic diagram of the ACTB gene is shown on top of the figure. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Antibody specificity was demonstrated by detection of enrichment of the targeted histone modification using SNAP-ChIP™ Spike-in, a proprietary technology developed by EpiCypher™. SNAP-ChIP™ spike-in was performed using H4K8ac polyclonal antibody (Product # PA5-16182) and H4K8ac was enriched compared to the other histone modifications in the SNAP-ChIP™ K-AcylStat Panel (Product # A47358).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Antibody specificity for modified targets can be established using peptide arrays by quantifying detection of the target protein along with closely related proteins. Peptide array of Histone H4K8ac using Anti-Acetyl-Histone H4 (Lys8) Antibody: An array of the specific peptide and other relevant peptides when tested using Anti-Acetyl-Histone H4 (Lys8) Polyclonal Antibody (Product # PA5-16182), showed that the Histone H4K8ac modification was specifically recognized by the antibody.
