Key features and details
- Mouse monoclonal [195C] to Werner's syndrome helicase WRN
- Suitable for: WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-Werner's syndrome helicase WRN antibody [195C]
See all Werner's syndrome helicase WRN primary antibodies
DescriptionMouse monoclonal [195C] to Werner's syndrome helicase WRN
Tested applicationsSuitable for: WB, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Human
Recombinant fragment within Human Werner's syndrome helicase WRN aa 1050 to the C-terminus. The exact sequence is proprietary. GST-WRN recombinant protein
Database link: Q14191
- WB: HAP1, MOLT-4 and K562 cells. IHC-P: Human testis seminoma tissue. ICC/IF: A431 cells
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact firstname.lastname@example.org.
Storage bufferPreservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
PurityProtein G purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab241545 in the following tested applications.
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 162 kDa.|
|ICC/IF||Use a concentration of 5 µg/ml.
4% PFA fixation for 10 minutes.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionMultifunctional enzyme that has both magnesium and ATP-dependent DNA-helicase activity and 3'->5' exonuclease activity towards double-stranded DNA with a 5'-overhang. Has no nuclease activity towards single-stranded DNA or blunt-ended double-stranded DNA. Binds preferentially to DNA substrates containing alternate secondary structures, such as replication forks and Holliday junctions. May play an important role in the dissociation of joint DNA molecules that can arise as products of homologous recombination, at stalled replication forks or during DNA repair. Alleviates stalling of DNA polymerases at the site of DNA lesions. Important for genomic integrity. Plays a role in the formation of DNA replication focal centers; stably associates with foci elements generating binding sites for RP-A.
Involvement in diseaseDefects in WRN are a cause of Werner syndrome (WRN) [MIM:277700]. WRN is a rare autosomal recessive progeroid syndrome characterized by the premature onset of multiple age-related disorders, including atherosclerosis, cancer, non-insulin-dependent diabetes mellitus, ocular cataracts and osteoporosis. The major cause of death, at a median age of 47, is myocardial infarction. Currently all known WS mutations produces prematurely terminated proteins.
Defects in WRN may be a cause of colorectal cancer (CRC) [MIM:114500].
Sequence similaritiesBelongs to the helicase family. RecQ subfamily.
Contains 1 3'-5' exonuclease domain.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain.
Contains 1 HRDC domain.
modificationsPhosphorylated by PRKDC. Phosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationNucleus > nucleolus. Nucleus.
- Information by UniProt
- DKFZp686C2056 antibody
- DNA helicase antibody
- DNA helicase, RecQ like type 3 antibody
All lanes : Anti-Werner's syndrome helicase WRN antibody [195C] (ab241545) at 1 µg/ml
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : WRN knockout HAP1 cell lysate
Lane 3 : MOLT-4 cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 162 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green - ab241545 observed at 170 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab241545 was shown to react with Werner's syndrome helicase WRN in wild-type HAP1 cells in western blot. Loss of signal was observed when WRN knockout sample was used. Wild-type and WRN knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab241545 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
IHC image of Werner's syndrome helicase WRN staining in a section of formalin-fixed paraffin-embedded human testis seminoma* performed on a Leica BOND™ 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 20mins. The section was then incubated with ab241545, 1μg/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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
ab241545 staining Werner's syndrome helicase WRN in A431 cells. The cells were fixed with 4% PFA (10mins), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Triton for 1h. The cells were then incubated overnight at +4°C with ab241545 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor®488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor®594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab241545 has not yet been referenced specifically in any publications.