Anti-phospho-Histone H2A.X (Ser139) Antibody, clone 6L16, ZooMAb® Rabbit Monoclonal
SIGMA/ZRB05636 - recombinant, expressed in HEK 293 cells
Synonym: H2a/x; Histone H2A.X
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human testis (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous/nuclear staining in human testis tissue sections was observed.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human testis (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous/nuclear staining in human testis tissue sections was observed.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human testis (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16, ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous/nuclear staining in human testis tissue sections was observed.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16 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 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16 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 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16 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 staurosporin was performed using a 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16 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 staurosporin was performed using a 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16 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 staurosporin was performed using a 1:100 dilution of Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16 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 Lysates from untreated Jurkat cells (Lane 1) and those treated with staurosporin (0.5 μM for 6 hours post serum starvation) (Lane 2) were probed with Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16 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 phospho-Histone H2A.X (Ser139) (~15 kDa).
Peptide Inhibition Enhanced Validation-Recombinant Antibody Technology Lysates from Jurkat cell lysates treated with staurosporin (0.5 μM 6 hours post serum starvation) were probed with Cat. No. ZRB05636, Anti-phospho-Histone H2A.X (Ser139), clone 6L16, ZooMAb® Rabbit Monoclonal each tested with a 1:1,000 dilution of antibody that was incubated with the following: Lane 1: No peptide added Lane 2: Histone H2Ax peptide without phosphorylation at serine residue 139 Lane 3: Histone H2Ax peptide with phosphorylation at serine residue 139 Image is a 5 minute exposure using crescendo.
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 | 6L16, 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 15.15 kDa |
| observed mol wt ~15 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 |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: suitable using 1:100 |
| 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. | P16104  |
| Application: | Anti-phospho-Histone H2A.X (Ser139), clone 6L16 ZooMAb, is a recombinant Rabbit Monoclonal Antibody that specifically targets phospho Histone H2AX (ser139) and has been tested for use in Immunocytochemistry, Immunohistochemistry (Paraffin), Peptide Inhibition Assay, and Western Blotting. |
| Application: | Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected phospho-Histone H2A.X (Ser139) in human testis tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-Histone H2A.X (Ser139) in HeLa treated with Staurosporin. Peptide Inhibition Analysis: A 1:1,000 dilution from a representative lot was used with Jurkat cell lysates treated with staurosporin 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 9 amino acids surrounding phospho-serine 139 of human Histone H2A.X. |
| 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: | 30 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 6L16 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects Histone H2A.X phosphorylated on serine 139. |
| 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 H2AX (UniProt: P16104; also known as H2a/x, Histone H2A.X) is encoded by the H2AFX (also known as H2AX) gene (Gene ID: 3014) 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. The histone H2A.X is a variant member of the H2A family of histones that is distinguished from other H2A histones by a unique carboxy-terminal sequence. This unique sequence is highly conserved throughout eukaryotic evolution and is rapidly phosphorylated by ATM or ATR at Serine 139 in mammals in response to DNA double-strand breaks. H2A.X phosphorylation is important in the formation of a stable repair complex at the site of DNA damage. Phosphorylation of H2A.X is important in the formation of a stable repair complex at the site of DNA damage. This phosphorylation is a very rapid response to DNA damage, occurring within as little as one minute after exposure to ionizing radiation. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |