Loading...
You have changed your country from
to
. Please be aware that this will change the currency in the purchasing process.
Products:Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> DNA Ligases
Overview
Product name
Anti-DNA Ligase IV antibody
See all DNA Ligase IV products (3) ...
Description
Rabbit polyclonal to DNA Ligase IV
Tested applications
WB, ICC/IF, IHC-Pmore details
Cross reactivity
Reacts with
Mouse, Human
Predicted to work with
Orangutan
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Human DNA Ligase IV.
(Peptide available as ab90599.)
Positive control
This antibody gave a positive signal in human testis tissue lysate and HepG2 whole cell lysate.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Synthesis >> DNA Ligases
Applications
Show applications key
Our Abpromise guarantee covers the use of ab80514 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).
ICC/IF: Use a concentration of 1 µg/ml
IHC-P: Use a concentration of 5 µg/ml
Target
Function
Efficiently joins single-strand breaks in a double-stranded polydeoxynucleotide in an ATP-dependent reaction. Involved in DNA non-homologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination. The LIG4-XRCC4 complex is responsible for the NHEJ ligation step, and XRCC4 enhances the joining activity of LIG4. Binding of the LIG4-XRCC4 complex to DNA ends is dependent on the assembly of the DNA-dependent protein kinase complex DNA-PK to these DNA ends.
Tissue specificity
Testis, thymus, prostate and heart.
Involvement in disease
Defects in LIG4 are the cause of LIG4 syndrome (LIG4S) [MIM:606593]. This disease is characterized by immunodeficiency and developmental and growth delay. Patients display unusual facial features, microcephaly, growth and/or developmental delay, pancytopenia, and various skin abnormalities.
Defects in LIG4 are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-negative/NK-cell-positive with sensitivity to ionizing radiation (RSSCID) [MIM:602450]. SCID refers to a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients with SCID present in infancy with recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development. Individuals affected by RS-SCID show defects in the DNA repair machinery necessary for coding joint formation and the completion of V(D)J recombination. A subset of cells from such patients show increased radiosensitivity.
Sequence similarities
Belongs to the ATP-dependent DNA ligase family.
Contains 2 BRCT domains.
Cellular localization
Nucleus.
Target information above from: UniProt accessionP49917
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- DNA ligase IV ATP dependent antibody
- DNA joinase antibody
- DNA ligase 4 antibody
see all
Database links
- Entrez Gene: 3981 Human
- Entrez Gene: 319583 Mouse
- Omim: 601837 Human
- SwissProt: P49917 Human
- SwissProt: Q8BTF7 Mouse
- Unigene: 166091 Human
- Unigene: 80584 Mouse
Anti-DNA Ligase IV antibody images:
Western blot - DNA Ligase IV antibody (ab80514)
All lanes : Anti-DNA Ligase IV antibody (ab80514) at 1 µg/ml
Lane 1 : Testis (Human) Tissue Lysate - adult normal tissue (ab30257)
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 103 kDa
Observed band size : 103 kDa
Additional bands at : 18 kDa,78 kDa,87 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 20 minutes
Immunocytochemistry/ Immunofluorescence - DNA Ligase IV antibody (ab80514)
ICC/IF image of ab80514 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 (ab80514, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit 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) - DNA Ligase IV antibody (ab80514)
IHC image of ab80514 staining in Human Normal Cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab80514, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
References for Anti-DNA Ligase IV antibody (ab80514)
This product has been referenced in:
- Gao Yet al. DNA ligase III is critical for mtDNA integrity but not Xrcc1-mediated nuclear DNA repair. Nature 471:240-4 (2011). WB; Mouse.Read more (PubMed: 21390131) »
See 1 publication for this product
Publishing research using ab80514? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"