Anti-Acetyl Histone H3 (Lys9 and 14) Antibody, clone 5H20, ZooMAb® Rabbit Monoclonal
SIGMA/ZRB06599 - recombinant, expressed in HEK 293 cells
Synonym: 14; 14Ac; H3/g; H3/t; H3K9; H3t; Histone H3 Ac Lysine 9; Histone H3.1t
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human breast cancer (Right issue) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Nuclear staining in human breast cancer tissue sections was observed.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human breast cancer (Right issue) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Nuclear staining in human breast cancer tissue sections was observed.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human breast cancer (Right issue) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Nuclear staining in human breast cancer tissue sections was observed.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells treated with Trichostatin A (TSA) was performed using a 1:1,000 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells treated with Trichostatin A (TSA) was performed using a 1:1,000 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells treated with Trichostatin A (TSA) was performed using a 1:1,000 dilution of Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Acid extract of HeLa cells treated with Sodium Butyrate (5 mM) was probed with Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14), clone 5H20, ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Acetyl Histone H3 (Lys9 and 14) (~16 kDa).
Peptide Inhibition Enhanced Validation-Recombinant Antibody Technology Acid extract of HeLa cells treated with 5 mM Sodium Butyrate (24 hours) was prepared. Blots were probed with Cat. No. ZRB06599, Anti-Acetyl Histone H3 (Lys9 and 14) clone 5H20, ZooMAb® Rabbit Monoclonal using a 1:1,000 dilution of antibody pre-mixed with the following peptides: Lane 1: No peptide Lane 2: Peptide with no acetylation modifications at Lysine 9 or 14 Lane 3: Peptide with acetylation modifications at Lysine 9 and 14 Image is a 2 minute exposure using forte.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit (recombinant) |
| clone | 5H20, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 15.51 kDa |
| observed mol wt ~16 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | mouse, rat |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: 1:100-1:1,000 |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable using 1:100 | |
| inhibition assay: suitable using 1:1,000 (peptide) | |
| western blot: suitable using 1:1,000 | |
| UniProt accession no. | Q16695  |
| Application: | Anti-Acetyl Histone H3 (Lys9 and 14), clone 5H20, ZooMAb, Cat. No. ZRB06599, is a recombinant Rabbit Monoclonal Antibody that specifically targets Histone H3 acetylated on Lysines 9 and 14 and has been tested for use in Immunocytochemistry, Immunohistochemistry (Paraffin), Peptide Inhibition, & WB. |
| Application: | Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected Acetyl Histone H3 (Lys9 and 14) in HeLa cells treated with Trichostatin A (TSA). Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Acetyl Histone H3 (Lys9 and 14) in human breast cancer tissue sections. Peptide Inhibition Analysis: A 1:1,000 dilution from a representative lot was used with HeLa cells treated with Sodium Butyrate for peptide block analysis. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to the first 21 amino acids (without methionine) surrounding acetylated lysines 9 and 14 of human Histone H3. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 5H20 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects Histone H3 acetylated on lysines 9 and 14. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Histone H3.1t (UniProt: Q16695; also known as H3/t, H3t, H3/g) is encoded by the HIST3H3 (also known as H3FT) gene (Gene ID: 8290) in human. Histones are highly conserved basic nuclear proteins that are responsible for the nucleosome structure of chromatin in eukaryotes. They play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which DNA is wrapped in repeating units, called nucleosomes, which limits DNA accessibility to the cellular machineries that require DNA as a template. Histone H3 features a main globular domain and a long N-terminal tail, which protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. Acetylation of histone H3 occurs at several different lysine positions in the histone tail and is performed by a family histone acetyl transferases. Histone H3 can also undergo methylation as well as acetylation on lysine 9. Acetylation on lysine 9 (H3K9ac) is reported to impair methylation at arginine 9 (H3R8me2s). A balance between acetylation and methylation is important for the establishment of specific chromatin structures. Acetylation of lysine 14 (H3K14Ac) is commonly seen in genes that are being actively transcribed into RNA. Histone H3 is reported to be hyperacetylated at lysines 9 and 14 following UV irradiation. Studies have shown that nucleosomes with H3K14ac modification have a higher affinity for purified chromatin remodeling complex RSC (Remodels the Structure of Chromatin) and by anchoring RSC, H3K14Ac plays an important role in the unfolding of strongly positioned nucleosomes during repair of UV damage. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Nicolas, E., et al. (2003). Mol. Cell. Biol. 23(5); 1614-1622; Duan, MR., and Smerdon, MJ (2014). J. Biol. Chem. 289(12); 8353-8363). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |