710828-20UG
antibody from Invitrogen Antibodies
Targeting: H4C14
H4/n, H4F2, H4FN, HIST2H4, HIST2H4A
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Chromatin Immunoprecipitation [1]
- Other assay [2]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 710828-20UG - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- H4K8ac Recombinant Polyclonal Antibody (9HCLC), ChIP-Verified
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
- Reactivity
- Human, Yeast
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 9HCLC
- Vial size
- 20 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A431 (Lane1), A431 treated with 0.5 uM SAHA for 24 hours (Lane 2), HeLa (Lane 3), HeLa treated with 0.5 uM SAHA for 24 hours (Lane 4), HeL 92.1.7 treated with 0.5 uM SAHA for 24 hours (Lane 5), HT 29 treated with 0.5 uM SAHA for 24 hours (Lane 6), HeLa treated with 5 mM sodium butyrate (Lane 7). The blots were probed with Anti-Histone H4K8ac Recombinant Rabbit Polyclonal Antibody (Product # 710828, 0.5-1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A clear 17kDa band corresponding to Histone H4K8ac was observed across cell lines according to the given treatment. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% nonfat milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of H4K8ac was performed by loading whole cell lysate in 1X SDS sample buffer with 2-ME from 4 yeast strains collected at a concentration of OD600= 10. Samples (30 µL) were loaded onto a 15% polyacrylamide gel. Proteins were transferred to nitrocellulose membrane with wet/tank transfer. Membrane was blocked in 5% milk/TBST. The membrane was probed with a recombinant polyclonal anti-H4K8ac antibody (Product # 710828-20UG) at a dilution of 1:500 in 5% milk/TBST at 4°C overnight, followed by a secondary antibody HRP-anti-rabbit at a dilution of 1:5,000 at room temperature for 1 hour. Chemiluminescent detection was performed. Data courtesy of the Antibody Data Exchange Program.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence was performed on fixed and permeabilized A549 cells for detection of Histone H4K8ac using Anti- Histone H4K8ac Recombinant Rabbit Polyclonal Antibody (Product # 710828, 1 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 0.4 µg/mL, 1:2500). Panel a) shows representative cells that were stained for detection and localization of Histone H4K8ac protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938, 1:50). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 594 Phalloidin (Product # A12381, 1:200). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Histone H4K8Ac. Panel e) represents control cells with no primary Antibody to assess background.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) was performed using Anti-Histone H4K8ac Recombinant Rabbit Polyclonal Antibody (Product # 710828, 5 µg) on sheared chromatin from 2 million Jurkat cells treated with 5mM sodium butyrate for 24 hours 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 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the promoters of the active cFOS, beta-actin genes, and IRF1 Exon 2 region used as positive control target gene, and the region of the inactive SAT2 satellite repeat, used as negative control target gene. 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 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) Recombinant Rabbit Polyclonal Antibody: An array of the specific peptide and other relevant peptides when tested using Anti-Acetyl-Histone H4 (Lys8) Recombinant Rabbit Polyclonal Antibody (Product # 710828), showed that the Histone H4K8ac modification was specifically recognized by the antibody.
- 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 Recombinant Rabbit Polyclonal Antibody (Product # 710828) and H4K8ac was enriched compared to the other histone modifications in the SNAP-ChIP™ K-AcylStat Panel (Product # A47358).