Anti-Werner's syndrome helicase WRN antibody [195C]

3 product images

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  Western blot - Anti-Werner's syndrome helicase WRN antibody [195C] (ab241545)

  Lanes 1 - 4: Merged signal (red and green). Green - ab241545 observed at 170 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - 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 Anti-GAPDH antibody [EPR16891] - Loading Control 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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Werner's syndrome helicase WRN antibody [195C] (ab241545) at 1 µg/mL

  Lane 1: Wild-type HAP1 cell lysate at 20 µg

  Lane 2: WRN knockout HAP1 cell lysate at 20 µg

  Lane 3: MOLT-4 cell lysate at 20 µg

  Lane 4: K562 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 162 kDa

  Observed band size: 170 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Werner's syndrome helicase WRN antibody [195C] (ab241545)

  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.

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  Immunocytochemistry/ Immunofluorescence - Anti-Werner's syndrome helicase WRN antibody [195C] (ab241545)

  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 Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed 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).