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Neutrophil Elastase protein (Active) (ab91099)

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Overview

  • Product nameNeutrophil Elastase protein (Active)See all Neutrophil Elastase proteins and peptides ...
  • Protein descriptionFull length native Human Neutrophil Elastase purified from Human Neutrophils; MWt 29.5kDa (UniProt P08246).
  • Expression hostNative

    • Properties

  • Purity> 95 % by SDS-PAGE
  • Biological activityActivity: 20-22 units per mg protein. One unit is defined as the amount of enzyme that hydrolyzes one micromole of Meo-suc-ala-pro-val-pNA per minute at 25°C in 100 mM Tris-HCl, pH 7.5, with 500 mM NaCl.
  • FormLyophilised
  • Storage instructionsShipped at 4°C. Store at -20ºC.This product is an active protein and may elicit a biological response in vivo, handle with caution.
  • Storage bufferPreservative: None
    Constituents: ddH20
  • Concentration information loading...
  • Additional notesReconstitute in 50 mM Na acetate pH 5.5 with 150 mM NaCl. Dissolve at a concentration of 1 to 2 mg/mL and store at -20 degrees C. If not immediately frozen, this enzyme should be used promptly to limit loss of activity. We do not suggest repeated thawing/freezing cycles. Protein Determination: Extinction Coefficient (E) 0.1% at 280nm, 1cm pathway = 0.985. Prepared from whole blood shown to be non reactive for HBsAg, anti-HCV, anti- HBc, and negative for anti-HIV 1 & 2 by FDA approved tests.
  • Research Areas

    • Applications

    Our Abpromise guarantee covers the use of ab91099 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Notes
    WB WB: Use at an assay dependent concentration. Ab91099 can be used as a WB positive control in conjunction with ab68672.
    SDS-PAGE SDS-PAGE: Use at an assay dependent dilution.
    Functional Studies FuncS: Use at an assay dependent dilution.

    Protein info

    • Alternative names
        Bone marrow serine proteaseELA2ELANE
        Elastase 2Elastase 2 neutrophilElastase neutrophil expressedElastase-2ELNE_HUMANGEGranulocyte derived elastaseHLEHNEHuman leukocyte elastaseLeukocyte elastaseMedullasinNENeutrophil elastasePMN EPMN elastasePolymorphonuclear elastaseSCN1
      see all
  • FunctionModifies the functions of natural killer cells, monocytes and granulocytes. Inhibits C5a-dependent neutrophil enzyme release and chemotaxis.
  • Tissue specificityBone marrow cells.
  • Involvement in diseaseDefects in ELANE are a cause of cyclic haematopoiesis (CH) [MIM:162800]; also known as cyclic neutropenia. CH is an autosomal dominant disease in which blood-cell production from the bone marrow oscillates with 21-day periodicity. Circulating neutrophils vary between almost normal numbers and zero. During intervals of neutropenia, affected individuals are at risk for opportunistic infection. Monocytes, platelets, lymphocytes and reticulocytes also cycle with the same frequency.
    Defects in ELANE are the cause of neutropenia severe congenital autosomal dominant type 1 (SCN1) [MIM:202700]. SCN1 is a disorder of hematopoiesis characterized by a maturation arrest of granulopoiesis at the level of promyelocytes with peripheral blood absolute neutrophil counts below 0.5 x 10(9)/l and early onset of severe bacterial infections.
  • Sequence similaritiesBelongs to the peptidase S1 family. Elastase subfamily.
    Contains 1 peptidase S1 domain.

    • Target information above from: UniProt accession P08246 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt

    Neutrophil Elastase protein (Active) images

    • SDS-PAGE - Neutrophil Elastase protein (Active) (ab91099)
      SDS-PAGE - Neutrophil Elastase protein (Active) (ab91099)
      SDS-PAGE: 12% Bis-Tris NuPAGE gel
      Lane 1: 5 µg Neutrophil Elastase (reduced/heated)
      Lane 2: 10 µg Neutrophil Elastase (reduced/heated)
      Lane 3: 20 µg Neutrophil Elastase (reduced/heated)
      Lane 4: Molecular weight markers
    • Western blot - Anti-Neutrophil Elastase antibody (ab68672)
      Western blot - Anti-Neutrophil Elastase antibody (ab68672)
      Anti-Neutrophil Elastase antibody (ab68672) at 1 µg/ml + Neutrophil Elastase protein (Active) (ab91099) at 0.01 µg

      Secondary
      Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
      developed using the ECL technique

      Performed under reducing conditions.

      Exposure time : 2 minutes

    References for Neutrophil Elastase protein (Active) (ab91099)

    ab91099 has not yet been referenced specifically in any publications.

    Publishing research using ab91099 ? Please let us know so that we can cite the reference in this datasheet

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    Submit an Abreview for ab91099 Submit a Question for ab91099

    Displaying 1 - 5 of 5 results for Abreviews and Q&A

    10
    Votes: Highest First

    Dear xxxxx,
    Many thanks for your help.
    We will readjust our Western protocol accordingly.
    Best wishes,

    Thats no problem. If you have any further questions, please do not hesitate to contact us again.

    Thank you for your reply but surely if there are 2 bands when you add more protein that would suggest that there is contamination with another protein other than the neutrophil elastase.
    Also adding 0.01ug is very low and I can still see 2 bands on t...

    Read More

    I now have some further information to explain the 14 kDa band observed in both the SDS page gel and Western blot of Neutrophil Elastase protein (Active), ab91099.

    I have been informed by the lab that performing SDS-page under reducing conditio...

    Read More

    Thank you for your reply but surely if there are 2 bands when you add more protein that would suggest that there is contamination with another protein other than the neutrophil elastase.
    Also adding 0.01ug is very low and I can still see 2 bands on t...

    Read More

    I understand your concerns in regards to this second band. From the SDS-page gel, it appears that the band at ˜14 kDa also increases with increasing concentration (it is quite low in the 5 ug lane). It therefore appears that the band is concentration depe...

    Read More

    Hello,
    Here is the additional information you asked for:
    1. What antibody have you been using to detect the Neutrophil
    Elastase protein?
    The antibody used in the two images that were forwarded to you was
    Monoclonal primary NE Ab [...

    Read More

    Thank you for providing me with that extra information.
    I now have some more information to share with you. It seems that the unexpected band at 14 kDa is more likely to be observed in the western blot if the sample is overloaded with protein. As can...

    Read More

    Good Morning,
    Following on with the issue described below, please find attached the
    images obtained with using the Abcam neutrophil elastase protein
    (ab91099) as a positive control in western blots.
    Hope you can help with its interpret...

    Read More

    Thank you for providing that information. I now understand more fully what your concerns regarding the protein are.
    It seems that the lower band is much more prominent (in comparison to the expected ˜25 kDa band) when blocked with BSA and a higher co...

    Read More

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