Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Flow cytometry [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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- Product number
- 720087 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- H4K20ac Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- These Polyclonal antibodies are of rabbit origin developed by immunizing animals with proteins or peptides. The polyclonal antibody is purified by affinity purification from the rabbit sera generated after immunizing the rabbits with a specific type of protein or peptide. The purified antibody is tested for its functionality in various relevant research applications. The antibody is developed for Research Use Only and is non-hazardous or non-infectious in nature.
- Concentration
- 0.5 mg/mL
Submitted references Deacetylation of Histone H4 Accompanying Cardiomyogenesis is Weakened in HDAC1-Depleted ES Cells.
Arcidiacono OA, Krejčí J, Suchánková J, Bártová E
International journal of molecular sciences 2018 Aug 16;19(8)
International journal of molecular sciences 2018 Aug 16;19(8)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on nuclear enriched cell extracts (30 µg lysate) of HCT116 (Lane1), HCT116 treated with Sodium butyrate (5mM/24 hours) (Lane 2), Jurkat (Lane 3), Jurkat treated with SAHA (0.5 uM/24 hours) (Lane 4) and Jurkat treated with Sodium butyrate (5mM/24 hours) (Lane 5). The blot was probed with Anti-Histone H4K20ac Rabbit Polyclonal Antibody (Product # 720087, 1-2 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 H4K20ac was observed across cell lines tested. 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% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence was performed on fixed and permeabilized A549 cells for detection of Histone H4K20ac using Anti-Histone H4K20ac Rabbit Polyclonal Antibody (Product # 720087, 0.5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Histone H4K20ac protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938,). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Histone H4K20ac. Panel e) represents control cells with no primary Antibody to assess background.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometry analysis of Histone H4K20ac was performed on HeLa cells treated with SAHA (0.5uM/16 hours) labeled with Anti-Histone H4K20ac Rabbit Polyclonal Antibody (Product# 720087, 2-4 ug/ 1M cells) or with rabbit isotype control and detected with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, (Alexa Fluor® 488 conjugate, Product # A27034, 0.4 ug/ml, 1:2500) as represented by the red and pink histograms respectively. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control. A representative of 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer (4468770).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) was performed using Anti-Histone H4K20ac Rabbit Polyclonal Antibody (Product # 720087, 5 ug) on sheared chromatin from 2 million HeLa 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 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
- Figure 3 Histone acetylation and methylation in HDAC1 wt and HDAC1 dn mESCs induced into cardiomyocytes and treated with HDACi. The level of H3K9ac, H3K9me3, H4ac, H4K20ac, pan-acetylated lysines (K-ac), and alpha-actinin in ( A ) HDAC1 wt mESCs and ( B ) HDAC1 dn mESCs. In three biological replicates, Western blots were performed on one gel. For the data presented in panel A or B, the gel was separated by Photoshop to show samples that were compared in one relevant subset. Data on histone levels were normalized to the level of histone H3 and non-histone proteins were normalized and quantified to the level of GAPDH ( C ). In wt and HDAC1 dn non-treated cells and in TSA-, SAHA-, or VPA-treated mESCs, panel ( Ca ) shows the levels of H4ac, ( Cb ) shows H4K20ac, and ( Cc ) shows the levels of alpha-actinin. The total protein levels were measured using a uQuant spectrophotometer for each sample, and an identical protein amount was loaded on the gels. In panel ( A , B ), the levels of histone markers are also shown for embryonic hearts (e15). Quantification of the protein levels in panel ( C ) was performed using ImageJ software (NIH, freeware). Statistical analyses were performed using Student's t -test; asterisks (*) in panel ( Ca - c ) show statistically significant differences at p