Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Neutrophil Elastase.
(Peptide available as ab68671.)
Our Abpromise guarantee covers the use of ab68672 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/200. (See abreview 24266)|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 29 kDa).Can be blocked with Human Neutrophil Elastase peptide (ab68671). Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.|
|IHC-Fr||1/100 - 1/250. PubMed: 22266908|
|IHC-P||1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Immunofluorescent staining with ab68672 of blood film of trangenic mouse that suffered from systemic inflamation. Neutrophil Elastase (in green) expressed in the lysosomes of activated neutrophils. Cells were fixed with methanol and permeabilized with 0.3% TritonX in 0.1% PBS. Blocking was performed with 10% serum for 15 minutes at 24ºC. This was followed by 1 hour incubation at 24ºC with ab68672 which was used at 1/200. The secondary antibody used was an anti-rabbit Alexa Fluor 488.
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
ab68672 staining Neutrophil Elastase in Mouse LPS treated Lung tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes. Samples were incubated with primary antibody (1/50) for 2 hours. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.
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).