ab131260 staining Neutrophil Elastase in Human bone marrow tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/5000). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

ab131260 staining Neutrophil Elastase in the HL-60 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/70). ab150120(1/500) an Alexa Fluor^®594-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.

Overlay histogram showing HL-60 cells stained with ab131260 (red line) at 1/20 dilution. The cells were fixed with 80% methanol. The secondary antibody used was a FITC conjugated goat anti-rabbit IgG at 1/150 dilution. Isotype control antibody (black line) was rabbit monoclonal IgG used under the same conditions. Cells also incubated without primary antibody and secondary antibody  (blue line)

ab131260 staining Neutrophil Elastase in Human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/5000). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

Immunohistochemical analysis of paraffin-embedded Human bone marrow tissue labelling Neutrophil Elastase with unpurified ab131260 at 1/250 dilution.

Anti-Neutrophil Elastase antibody [EPR7479] (ab131260) at 1/1000 dilution + HL-60 Cell Lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 29 kDa
Observed band size: 29 kDa

Overlay histogram showing HL60 cells stained with unpurified ab131260 (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 (ab131260, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor® 488 (IgG H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.

Anti-Neutrophil Elastase antibody [EPR7479] (ab131260) at 1/1000 dilution (Unpurified) + HL60 cell lysate at 10 µg

Secondary
HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 29 kDa

Secondary antibody - goat anti-rabbit HRP (ab6721)

Equilibrium disassociation constant (K_D)
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