Anti-Lamin A antibody [133A2] - Nuclear Envelope Marker

8 product images

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  Flow Cytometry - Anti-Lamin A antibody [133A2] (ab8980)

  Overlay histogram showing HeLa cells stained with ab8980 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8980, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG3 [MG3-35] (2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.
  
  

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  Western blot - Anti-Lamin A antibody [133A2] - Nuclear Envelope Marker (ab8980)

  Lamin A Western blot staining using mouse Anti-Lamin A antibody

  Lanes 1 - 4: Merged signal (red and green). Green - ab8980 observed at 74 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.
  

  ab8980 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in LMNA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and LMNA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3pc Milk. ab8980 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/100 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Lamin A antibody [133A2] - Nuclear Envelope Marker (ab8980) at 1/100 dilution

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

  Lane 2: LMNA knockout HAP1 whole cell lysate at 20 µg

  Lane 3: HeLa whole cell lysate at 20 µg

  Lane 4: HepG2 whole cell lysate at 20 µg

  Predicted band size: 74 kDa

  Observed band size: 74 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)

  ICC/IF image of ab8980 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum (Normal Goat Serum ab7481) / 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 ab8980 at 1μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) 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.

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  This image is courtesy of a customer review submitted by Mr Carl Hobbs

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)

  ab8980 staining of Lamin A in adult mouse liver sections. Formaldehyde-fixed paraffin-embedded sections of mouse liver tissue were incubated with ab8980 (1/2500) fo 2 hours. Antigen retrieval was performed by heat induction in citrate buffer pH 6.0 (10x Citrate Buffer pH 6.0 ab64214). Please see accompanying abreview for additional information.

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  Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)

  These images show HeLa cells stained with Laminin A antibody - ab8980 (green) and with DAPI (blue). The pictures were kindly supplied as part of the review submitted by Dr Josef Gotzmann at Medical University of Vienna, Austria.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)

  Immunohistochemistry on paraffin section of human colon.

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  Immunohistochemistry (Frozen sections) - Anti-Lamin A antibody [133A2] (ab8980)

  Immunohistochemistry on frozen sections of human colon showing nuclear lamina staining in epithelial and connective tissue cells.

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  Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)

  Immunocytochemical staining of fiboblasts showing nuclear lamina