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Product Info Summary
| SKU: | PB10058 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Mouse, Rat |
| Host: | Rabbit |
| Application: | IHC, WB |
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Product info
Product Name
Anti-Neutrophil Elastase/ELANE Antibody Picoband®
SKU/Catalog Number
PB10058
PB1114 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
ELANE (neutrophil elastase) is a serine protease released from neutrophil granules that contributes to antimicrobial defense and inflammatory tissue remodeling. Assay context: IHC for neutrophil-rich lesions; Western blot for lysate confirmation. Commonly analyzed together with oxidative degranulation markers such as MPO when quantifying neutrophil activation states across tissues/conditions.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Neutrophil Elastase/ELANE Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB10058)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived mouse Elastase/ELANE/ELA2 recombinant protein (Position: S27-N265). Mouse Elastase/ELANE/ELA2 shares 92% amino acid (aa) sequence identity with rat Elastase/ELANE/ELA2.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB10058 is reactive to Elane in Mouse, Rat
Observed Molecular Weight
29 kDa
Calculated molecular weight
28.6 kDa
Background of Elane
Neutrophil elastase is a serine protease of neutrophil and monocyte granules. Its key physiologic role is in innate host defense, but it can also participate in tissue remodeling and possesses secretagogue actions important to local inflammatory responses. Elastases form a subfamily of serine proteases that hydrolyze many proteins in addition to elastin. Humans have six elastase genes which encode structurally similar proteins. The encoded preproprotein is proteolytically processed to generate the active protease. Following activation, this protease hydrolyzes proteins within specialized neutrophil lysosomes, called azurophil granules, as well as proteins of the extracellular matrix. This protein also degrades the outer membrane protein A (OmpA) of E. coli as well as the virulence factors of such bacteria as Shigella, Salmonella and Yersinia. Mutations in this gene are associated with cyclic neutropenia and severe congenital neutropenia (SCN). This gene is present in a gene cluster on chromosome 19.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
PB10058 is guaranteed for IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Mouse
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat
ELISA, 0.1-0.5μg/ml,-
Positive Control
WB: mouse spleen tissue
IHC: mouse spleen tissue, rat spleen tissue
Validation Images & Assay Conditions
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Western blot analysis of Elastase/ELANE/ELA2 using anti-Elastase/ELANE/ELA2 antibody (PB10058).
Electrophoresis was performed on a 13% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: mouse spleen tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Elastase/ELANE/ELA2 antigen affinity purified polyclonal antibody (PB10058) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Elastase/ELANE/ELA2 at approximately 29 kDa. The expected band size for Elastase/ELANE/ELA2 is at 29 kDa.
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IHC analysis of Elastase/ELANE/ELA2 using anti-Elastase/ELANE/ELA2 antibody (PB10058).
Elastase/ELANE/ELA2 was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Elastase/ELANE/ELA2 Antibody (PB10058) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IHC analysis of Elastase/ELANE/ELA2 using anti-Elastase/ELANE/ELA2 antibody (PB10058).
Elastase/ELANE/ELA2 was detected in a paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Elastase/ELANE/ELA2 Antibody (PB10058) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Neutrophil infiltration and NETs release within AAA lesions. A IF staining for neutrophil marker (Ly6G) was performed on tissue sections of the mouse abdominal aorta. B Hyalinization and IF labeling of the whole abdominal aorta and 3D tissue imaging were acquired with light-sheet microscopy. C – E Quantification of NETs markers (ELANE and MPO) in mouse abdominal aorta by western blotting ( n = 3). ** P < 0.01, **** P < 0.0001, indicating statistical significance between the two groups.
Index in PubMed under a CC BY license. PMID: 39237520
Specific Publications For Anti-Neutrophil Elastase/ELANE Antibody Picoband® (PB10058)
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6 Customer Q&As for Anti-Neutrophil Elastase/ELANE Antibody Picoband®
Question
I have a question about product PB10058, anti-Neutrophil Elastase/ELANE antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
B. Jackson
Verified customer
Asked: 2018-06-26
Answer
It is not recommended storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free PB10058 anti-Neutrophil Elastase/ELANE antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2018-06-26
Question
Would anti-Neutrophil Elastase/ELANE antibody PB10058 work on canine WB with lung?
Verified Customer
Verified customer
Asked: 2018-03-28
Answer
Our lab technicians have not validated anti-Neutrophil Elastase/ELANE antibody PB10058 on canine. You can run a BLAST between canine and the immunogen sequence of anti-Neutrophil Elastase/ELANE antibody PB10058 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated canine samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in canine lung in WB, you can get your next antibody for free.
Boster Scientific Support
Answered: 2018-03-28
Question
We are currently using anti-Neutrophil Elastase/ELANE antibody PB10058 for rat tissue, and we are content with the WB results. The species of reactivity given in the datasheet says mouse, rat. Is it possible that the antibody can work on monkey tissues as well?
Verified Customer
Verified customer
Asked: 2018-01-26
Answer
The anti-Neutrophil Elastase/ELANE antibody (PB10058) has not been tested for cross reactivity specifically with monkey tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in monkey you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2018-01-26
Question
Would PB10058 anti-Neutrophil Elastase/ELANE antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2017-09-07
Answer
You can see on the product datasheet, PB10058 anti-Neutrophil Elastase/ELANE antibody as been validated on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2017-09-07
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for bone marrow cell using anti-Neutrophil Elastase/ELANE antibody PB10058. Let me know if you need anything else.
D. Anderson
Verified customer
Asked: 2014-03-12
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2014-03-12
Question
I see that the anti-Neutrophil Elastase/ELANE antibody PB10058 works with WB, what is the protocol used to produce the result images on the product page?
M. Singh
Verified customer
Asked: 2013-01-08
Answer
You can find protocols for WB on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to support@bosterbio.com
Boster Scientific Support
Answered: 2013-01-08
