![Immunocytochemistry/ Immunofluorescence - Ku70 antibody [N3H10] (ab3114) image](#Ku70-Primary-antibodies-ab3114-1.jpg)
 image](#Ku70-Primary-antibodies-ab3114-2.jpg)
![Western blot - Anti-Ku70 antibody [N3H10] (ab3114) image](#Ku70-Primary-antibodies-ab3114-22.jpg)
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Product name
Anti-Ku70 antibody [N3H10]
See all Ku70 products (22) ...
Description
Mouse monoclonal [N3H10] to Ku70
Tested applications
IF, IHC-P, IP, WB, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Human, Monkey
Predicted to work with
Hamster, Xenopus laevis
Does not react with
Rabbit, Cow, Sea Urchin
Immunogen
Human placental extract designated as PSE1-PL.
Epitope
Amino acids 506 - 541.
Positive control
BT474, HEP-G-2 or HeLa cells. Tonsil tissue sections.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
10mM PBS, pH7.4, 0.2%BSA, 0.09% sodium azide
Concentration
Concentration information loading...
Purity
Protein A purified
Clonality
Monoclonal
Clone number
N3H10
Isotype
IgG2b
Research areas
Cancer >> Cell cycle >> Cell division
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> Non Homol. End Joining
Applications
Show applications key
Our Abpromise guarantee covers the use of ab3114 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IF: Use at an assay dependent dilution.
IHC-P: Use a concentration of 1 - 2 µg/ml.
IP: Use at an assay dependent dilution. (with protein A): use at a concentration of 2µg/mg of protein lysate (native and denatured).
WB: Use a concentration of 0.25 - 0.5 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).
ICC/IF: Use a concentration of 1 µg/ml.
Target
Function
Single stranded DNA-dependent ATP-dependent helicase. Has a role in chromosome translocation. The DNA helicase II complex binds preferentially to fork-like ends of double-stranded DNA in a cell cycle-dependent manner. It works in the 3'-5' direction. Binding to DNA may be mediated by XRCC6. Involved in DNA non-homologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination. The XRCC5/6 dimer acts as regulatory subunit of the DNA-dependent protein kinase complex DNA-PK by increasing the affinity of the catalytic subunit PRKDC to DNA by 100-fold. The XRCC5/6 dimer is probably involved in stabilizing broken DNA ends and bringing them together. The assembly of the DNA-PK complex to DNA ends is required for the NHEJ ligation step. Required for osteocalcin gene expression. Probably also acts as a 5'-deoxyribose-5-phosphate lyase (5'-dRP lyase), by catalyzing the beta-elimination of the 5' deoxyribose-5-phosphate at an abasic site near double-strand breaks. 5'-dRP lyase activity allows to 'clean' the termini of abasic sites, a class of nucleotide damage commonly associated with strand breaks, before such broken ends can be joined. The XRCC5/6 dimer together with APEX1 acts as a negative regulator of transcription.
Sequence similarities
Belongs to the ku70 family.
Contains 1 Ku domain.
Contains 1 SAP domain.
Developmental stage
Expression does not increase during promyelocyte differentiation.
Post-translational
modifications
Phosphorylation by PRKDC may enhance helicase activity. Phosphorylation of Ser-51 does not affect DNA repair.
Cellular localization
Nucleus. Chromosome.
Target information above from: UniProt accessionP12956
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- 5''-deoxyribose-5-phosphate lyase Ku70 antibody
- 5''-dRP lyase Ku70 antibody
- 70 kDa subunit of Ku antigen antibody
see all
Database links
- Entrez Gene: 2547 Human
- Entrez Gene: 14375 Mouse
- Omim: 152690 Human
- SwissProt: P12956 Human
- SwissProt: P23475 Mouse
- Unigene: 292493 Human
- Unigene: 288809 Mouse
Anti-Ku70 antibody [N3H10] images:
Immunocytochemistry/ Immunofluorescence - Ku70 antibody [N3H10] (ab3114)
![Immunocytochemistry/ Immunofluorescence - Ku70 antibody [N3H10] (ab3114)](/ps/datasheet/images/3/ab3114/Ku70-Primary-antibodies-ab3114-1.jpg)
ICC/IF image of ab3114 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3114, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Ku70 antibody [N3H10](ab3114)
](/ps/datasheet/images/3/ab3114/Ku70-Primary-antibodies-ab3114-2.jpg)
Ab3114 staining human malignant breast carcinoma. Staining is localized to nuclear compartment.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Western blot - Anti-Ku70 antibody [N3H10] (ab3114)
![Western blot - Anti-Ku70 antibody [N3H10] (ab3114)](/ps/datasheet/images/3/ab3114/Ku70-Primary-antibodies-ab3114-22.jpg)
Anti-Ku70 antibody [N3H10] (ab3114) at 1/500 dilution + whole cell lysate prepared from HeLa cells at 10 µg
Secondary
HRP conjugated goat anti-mouse IgG at 1/5000 dilution
developed using the ECL technique
Predicted band size : 70 kDa
Exposure time : 1 minute
Cells were lysed in SDS sample buffer plus beta-mercaptoethanol.Gels were transferred to PVDF by semi-dry transfer.
Image courtesy of an anonymous Abreview.
References for Anti-Ku70 antibody [N3H10] (ab3114)
This product has been referenced in:
- Shang ZF et al. Inactivation of DNA-dependent protein kinase leads to spindle disruption and mitotic catastrophe with attenuated checkpoint protein 2 Phosphorylation in response to DNA damage. Cancer Res 70:3657-66 (2010). WB; Human.Read more (PubMed: 20406977) »
- Strang BL et al. Nucleolin associates with the human cytomegalovirus DNA polymerase accessory subunit UL44 and is necessary for efficient viral replication. J Virol 84:1771-84 (2010). WB; Human.Read more (PubMed: 20007282) »
See all 6 publications for this product
Publishing research using ab3114? Please let us know so that we can cite the reference in this datasheet
![Anti-Ku70 antibody [N3H10] for Western blot in Human (3114)](/ps/datasheet/images/3/ab3114/Ku70-Primary-antibodies-ab3114-11.jpg)
![Ku70 antibody [N3H10] for Western blot in Human (3114)](/ps/datasheet/images/3/ab3114/Ku70-Primary-antibodies-ab3114-4.jpg)
![Anti-Ku70 antibody [N3H10] for Immunocytochemistry/ Immunofluorescence in Human (3114)](/ps/datasheet/images/3/ab3114/Ku70-Primary-antibodies-ab3114-20.jpg)
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"