Anti-CLASP1 antibody [EPR3409] - BSA and Azide free

8 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling CLASP1 (green) with Anti-CLASP1 antibody [EPR3409] ab108620 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

  Control: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).

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  Western blot - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).
  

  Lanes 1 - 4: Merged signal (red and green). Green - Anti-CLASP1 antibody [EPR3409] ab108620 observed at 169 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.
  
  Anti-CLASP1 antibody [EPR3409] ab108620 was shown to recognize CLASP1 in wild-type HAP1 cells as signal was lost at the expected MW in CLASP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CLASP1 knockout samples were subjected to SDS-PAGE. Anti-CLASP1 antibody [EPR3409] ab108620 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576) at 1/10000 dilution

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

  Lane 2: CLASP1 knockout HAP1 whole cell lysate at 20 µg

  Lane 3: HeLa whole cell lysate at 20 µg

  Lane 4: Jurkat whole cell lysate at 20 µg

  Predicted band size: 169 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling CLASP1 with purified Anti-CLASP1 antibody [EPR3409] ab108620 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling CLASP1 with unpurified Anti-CLASP1 antibody [EPR3409] ab108620 at 1/250.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).

  Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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  Immunocytochemistry/ Immunofluorescence - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling CLASP1 with unpurified Anti-CLASP1 antibody [EPR3409] ab108620 at 1/100.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).

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  Flow Cytometry (Intracellular) - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  Intracellular Flow Cytometry analysis of HeLa cells labelling CLASP1 with purified Anti-CLASP1 antibody [EPR3409] ab108620 at 1/50 (red). Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labeling CLASP1 with purified Anti-CLASP1 antibody [EPR3409] ab108620 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).

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  Flow Cytometry (Intracellular) - Anti-CLASP1 antibody [EPR3409] - BSA and Azide free (ab232576)

  Overlay histogram showing HeLa cells stained with unpurified Anti-CLASP1 antibody [EPR3409] ab108620 (red line). The cells were fixed with 80% 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 (Anti-CLASP1 antibody [EPR3409] ab108620, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CLASP1 antibody [EPR3409] ab108620).