Anti-p-DNA-PKcs-Ser2056 Antibody, clone 1K3 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1703 - recombinant, expressed in HEK 293 cells
Synonym: DNA-dependent protein kinase catalytic subunit;EC:2.7.11.1;DNA-
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/nucleolar staining was observed in glandular cells & lamina propria of Human Colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/nucleolar staining was observed in glandular cells & lamina propria of Human Colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/nucleolar staining was observed in glandular cells & lamina propria of Human Colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/nucleolar staining was observed in glandular cells & lamina propria of Human Colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/nucleolar staining was observed in glandular cells & lamina propria of Human Colon tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated Jurkat cells (A and B) and Jurkat cells treated with camptothecin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 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 untreated Jurkat cells (A and B) and Jurkat cells treated with camptothecin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 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 untreated Jurkat cells (A and B) and Jurkat cells treated with camptothecin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 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 untreated Jurkat cells (A and B) and Jurkat cells treated with camptothecin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 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 untreated Jurkat cells (A and B) and Jurkat cells treated with camptothecin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 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 Jurkat cells treated with Campthothecin (1 mM; 3 h) (Lane 2) were probed with Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-DNA-PKcs-Ser2056 (~350 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or phospho-DNA PKcs-Ser2056 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal. The binding of this antibody to phospho-DNA PKcs-Ser2056 peptide displayed a KD of 9.3 x 10-7, whereas binding to non-specific control peptide displayed a KD of 1.7 x 10-3.
Peptide Inhibition Assay Enhanced Validation - Recombinant Antibody Technology Lysates from Jurkat treated with Campthothecin (1 mM; 3 h) were probed with Cat. No. ZRB1703, Anti-p-DNA-PKcs-Ser2056, clone 1K3 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-DNA-PKcs-Ser2056 (~350 kDa). Lanes 1: Incubated without any peptide. 2: Incubated with phosphorylated DNA PKcs-Ser2056 peptide. 3: Incubated with non-phosphorylated DNA PKcs-Ser2056 peptide.
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 |
| clone | 1K3, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Internal |
| 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 469.09 kDa |
| observed mol wt ~350 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_008835  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| peptide synthesis: suitable | |
| western blot: suitable | |
| UniProt accession no. | P78527 |
| Application: | Quality Control Testing Evaluated by Immunohistochemistry (Paraffin) in human colon tissue sections. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution of this antibody detected phospho-DNA-PKcs-Ser2056 in human colon tissue sections. Tested applications Peptide Inhibition Assay: Target band detection in lysates from Jurkat cells treated with 1 mM Camptothecin (3 h) was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide. Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected phospho-DNA-PKcs-Ser2056 in Jurkat cells treated with Campthothecin ( 1 mM; 3 h). Affinity Binding Assay: A representative lot of this antibody bound phospho-DNA PKcs-Ser2056 with a KD of 9.3 x 10-7 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-DNA-PKcs-Ser2056 in Jurkat cells treated with Camptothecin. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| 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. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 14 amino acids surrounding phosphoserine 2056 in the internal region of human DNA-dependent protein kinase catalytic subunit (DNS-PKcs). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. 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 per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1K3 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects DNA-dependent protein kinase catalytic subunit (DNA-PKcs) phosphorylated on serine 2056. |
| 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: | DNA-dependent protein kinase catalytic subunit (UniProt: P78527; also known as EC:2.7.11.1, DNA-PK catalytic subunit, DNA-PKcs, DNPK1, p460) is encoded by the PRKDC (also known as HYRC, HYRC1) gene (Gene ID: 5591) in human. DNA-PK is a heterotrimeric, nuclear serine/threonine-protein kinase that consists of PRKDC and the Ku p70/YRCC6-p86/XRCC5 dimer. It is involved in cell cycle control, DNA repair, and DNA damage responses. It serves as a molecular sensor for DNA damage and is involved in DNA non-homologous end joining required for double-strand break (DSB) repair and V(D)J recombination. It may also serve as a scaffold protein to aid the localization of DNA repair proteins to the site of damage. DNA-PK is also present at the ends of chromosomes, suggesting a role in the maintenance of telomeric stability and the prevention of chromosomal end fusion. Its catalytic activity is expressed upon binding to DNA. DNA-PKcs undergoes autophosphorylation at two clusters, the Thr2609 and the Ser2056, which are DNA damage-inducible phosphorylation sites. Autophosphorylation of the Thr2609 cluster is shown to be essential for hematopoietic development and protein synthesis in erythrocytes precursors. Autophosphorylation induces a conformational change that leads to remodeling of the DNA-PK complex, which is a requisite for efficient end processing and DNA repair. Although DNA-PKcs falls into the phosphatidylinositol (PI) 3-kinase family, it does not exhibit any no detectable activity toward lipids. Deficiency of DNA-PKcs is linked to severe combined immunodeficiency characterized by reduced or absent T and B cells, recurrent candidiasis, and lower respiratory tract infections. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Ma, Y., et al. (2004). Mol. Cell. 16(5); 701-713; Wechsler, T., et al. (2004). Proc. Nall. Acad. Sci. USA. 101(5); 1247-1252; Douglas, P., et al. (2002). Biochem. J. 368(1); 243-251). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |