Anti-Neutrophil Elastase antibody (ab68672)
Key features and details
- Rabbit polyclonal to Neutrophil Elastase
- Suitable for: WB, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Neutrophil Elastase antibody
See all Neutrophil Elastase primary antibodies -
Description
Rabbit polyclonal to Neutrophil Elastase -
Host species
Rabbit -
Specificity
We have had mixed results for use of this antibody in IHC-P using Mouse tissue samples. The abReview data below from external researchers reports good staining with this antibody in Mouse tissue samples using IHC-Fr. -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB MouseHuman
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Immunogen
Synthetic peptide corresponding to Human Neutrophil Elastase aa 1-100 (internal sequence) conjugated to keyhole limpet haemocyanin.
(Peptide available asab68671) -
Positive control
- WB: Mouse Bone Marrow, U-937 Whole Cell Lysate, Native human Neutrophil Elastase protein. IHC-P: Human spleen.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Images
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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
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Western blot - Anti-Neutrophil Elastase antibody (ab68672)All lanes : Anti-Neutrophil Elastase antibody (ab68672) at 1 µg/ml
Lane 1 : Bone Marrow (Human) Tissue Lysate - adult normal tissue at 20 µg
Lane 2 : Mouse Bone Marrow Tissue Lysate at 20 µg
Lane 3 : HepG2 Whole Cell Lysate (Negative Control) at 10 µg
Lane 4 : U-937 Whole Cell Lysate at 10 µg
Lane 5 : U-937 (PMA Treated) Whole Cell Lysate (Negative Control)
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 29 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab68672 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Neutrophil Elastase antibody (ab68672) This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United KingdomImmunohistochemistical 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).
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Western blot - Anti-Neutrophil Elastase antibody (ab68672)Anti-Neutrophil Elastase antibody (ab68672) at 1 µg/ml +Native human Neutrophil Elastase protein (Active) (ab91099) at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 29 kDa
Exposure time: 2 minutes
