Anti-Neutrophil Elastase antibody

• IHC-P

AbReview18305****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody (AB68672)

Immunohistochemistical detection of Neutrophil Elastase using antibody (ab68672) on formaldehyde-fixed paraffin-embedded human spleen sections. Antigen retrieval step : heat mediated in citric acid pH6 buffer. Permeabilization : No Blocking step : 1% BSA for 10 mins @ rt°C. Primary antibody dilution 1/2000; incubation time : 2 hours in TBS/BSA/azide. Secondary Antibody : anti Rabbit IgG conjugated to biotin (1/200).

This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody (AB68672)

IHC image of Neutrophil Elastase staining in human spleen formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system 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 20 mins. The section was then incubated with ab68672, 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.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody (AB68672)

Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling Neutrophil Elastase with ab68672 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

Anti-Neutrophil Elastase antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody (AB68672)

IHC image of Neutrophil Elastase staining in a section of formalin-fixed paraffin-embedded normal mouse spleen performed on a Leica Biosystems BOND® RX instrument. 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 ab68672, 5ug/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.

• WB

Unknown

Western blot - Anti-Neutrophil Elastase antibody (AB68672)

All lanes:

Western blot - Anti-Neutrophil Elastase antibody (ab68672) at 1 µg/mL

All lanes:

Western blot - Native human Neutrophil Elastase protein (Active) (<a href='/en-us/products/proteins-peptides/native-human-neutrophil-elastase-protein-active-ab91099'>ab91099</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 29 kDa

true

Exposure time: 2min

• IHC

CiteAb

Immunohistochemistry - Anti-Neutrophil Elastase antibody (AB68672)

Immunohistochemistry using Anti-Neutrophil Elastase antibody, ab68672. Publication image from Han, W. et al., 2016, Nat Commun, 27046438. Legend direct from paper.

Lung inflammation progresses with age in pIgR−/− mice.(a) Representative immunostains for neutrophils using antibodies to neutrophil elastase (NE) or macrophages using antibodies to CD68 in 12-month-old WT and pIgR−/− mice. Positive cells are stained brown (indicated by red arrows) (original magnification, x 200). Scale bar, 50 µm. (b–e) Neutrophil (NE+) and macrophage (CD68+) counts in lungs of pIgR−/− and age-matched WT littermate controls at the indicated ages, and neutrophil and macrophage counts in BAL fluid. Five to seven mice per group; *P<0.05 compared with 2-month-old pIgR−/− mice and age-matched WT mice; **P<0.01 compared with all other groups (two-way analysis of variance (ANOVA)). (f) Representative image of folate-PEG-Cy5-derived chest fluorescence 4 h after intravenous probe injection in 12-month-old WT and pIgR−/− mice. (g) Photon emission from the chest normalized to background before injection of probe. Three to four mice per group; *P<0.05 (Student's t-test). (h) Western blotting and densitometry for MMP-12 (two bands at 45 and 54 kDa) and NE (29 kDa) in lung tissue from 12-month-old WT and pIgR−/− mice. Band densities of MMP-12 and NE were normalized to β-actin. Six mice per group; *P<0.01 compared with WT mice (Student's t-test).