Overview

  • Product name
    Anti-Legionella pneumophila antibody (Biotin)
    See all Legionella pneumophila primary antibodies
  • Description
    Rabbit polyclonal to Legionella pneumophila (Biotin)
  • Host species
    Rabbit
  • Conjugation
    Biotin
  • Tested applications
    Suitable for: ICC/IF, ELISAmore details
  • Species reactivity
    Reacts Legionella pneumophila serogroups 1-12, other organisms not tested.
  • Immunogen

    A whole cell preparation of Legionella pneumophila; ATCC #33152

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituent: 0.0268% PBS
  • Concentration information loading...
  • Purity
    IgG fraction
  • Purification notes
    Purified IgG fraction covalently coupled with the N-Hydroxysuccinimide ester of biotin under mild conditions to give a high degree of substitution.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab20527 in the following tested applications.

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

Application Abreviews Notes
ICC/IF
ELISA
  • Application notes
    ELISA: Use at an assay dependent dilution.
    IF: Acetone-fixed organisms. 1/800.

    Not tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Relevance
      Legionella pneumophila is a flagellated gram negative bacterium found primarily in warm water environments. Legionella pneumophila causes a type of pneumonia called Legionnaire disease and a milder condition called Pontiac fever. Infection is acquired through inhalation of the microorganism in water mists (e.g. from air conditioning, cooling towers, whirlpool spas and showers mists). A number of risk factors for acquiring Legionnaire disease have been identified, including age, smoking, chronic lung disease, cancer, and immunosuppression.

    References

    ab20527 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Answer

    ab20561 and ab20527 are produced in the same lab and in exactly the same way, using a whole cell preparation of Legionella pneumophila; ATCC #33152 to immunise the rabbits. They are therefore the same antibody but conjugated to different molecules. ab20561 is HRP conjugated whereas ab20527 is biotin conjugated.

    Since these antibodies are polyclonal antibodies, they will recognise multiple epitopes on any one antigen which is different to monoclonal antibodies which detect only one epitope on the antigen. Therefore, the epitopes recognised by these antibodies will be batch dependent and so we cannot say whether these two antibodies will react against the same epitope.

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    Question

    We are developing a fast screening method for L. pneumophila based on immunostaining with FITC coupled surface antibody / immunomagnetic separation and flow cytometric detection. We got very good recovery rate but one problem is that background and positive signal are quite near together. Therefore we want to double stain or magnify the fluorescence of the FITC-labelled L. pneumophila cells. We label our cells with one of your FITC conjugated antibody for L. pneumophila ( Abcam ab20818). We are working with a Flow cytometer (Cyflow Partec) with three lasers ( UV(375): 25 mW, Blue laser ( 488 nm): 200 mW, Green Laser (525 nm): 25 mW). I worked out two possible solutions: 1. Signal Amplification (520 nm): Anti-FITC coupled with Alexa Fluor 488 conjugate Question: 1. Can you recommend an Anti-FITC antibody for amplifying the signal? 2. I would be also interested for Anti-FITC antibody conjugated with other stain wich can ecited by UV- / Blue- or Greenlaser and excite at 375 nm , 520 nm , 590 nm or 630 nm. 2. Doublestain a) Specific antibody for L. pneumophila with a conjugate which can be excited by UV- or Green laser. Question: 1. Can you recommend me an antibody for L. pneumophila which is conjugated with another stain than FITC? 2. With a Antibody labelling Kit from Invitrogen I can label unconjugated IgG antibody for L. pneumphila. I see that you have several unconjugated antibody for L. pneumophila. Which one do you recommend me. I suppose that if possible it would be better to use another antibody as used my first antibody (Abcam ab20818) in order to avoid/minimise a competition for free epitopes on the surface of L. pneumophila.

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    Answer

    Thank you for contacting us and for presenting your questions so clearly! To amplify the the signal from the FITC-conjugated anti-Legionella antibody, your idea of using an Alexa Fluor 488-conjugated anti-FITC is a good one. We unfortunately do not have Alexa Fluor conjugates but these dyes have a good reputation for strong, stable signal. Another option is to follow the primary antibody with a biotinylated anti-rabbit IgG such as ab6720, and then a streptavidin or avidin fluorochrome conjugate. Again, we do not have these streptavidin/avidin/fluorochrome conjugates but they are available from other companies. The anti-rabbit IgG secondary will bind ab20818 even though it has FITC conjugated to it: https://www.abcam.com/index.html?datasheet=6720 We do have a phycoerythrin-conjugated anti-FITC, ab25539: https://www.abcam.com/index.html?datasheet=25539 We do not have another fluorochrome-conjugated anti-Legionella antibody but we do have a biotin conjugate, ab20527: https://www.abcam.com/index.html?datasheet=20527 This can be used with the streptavidin/avidin conjugates I mentioned above. An antibody appropriate for labelling should have no other proteins in the buffer it is delivered in, as these will compete with the antibody for the label. Another consideration is that, since you already have a rabbit polyclonal antibody that will recognize potentially several epitopes, and it is not clear what exactly on the bacterium it binds, another polyclonal antibody applied before this one may interfere with it, and vice versa. Your best strategy, aside from using two monoclonal antibodies, is to apply a monoclonal antibody first, such as ab20444: https://www.abcam.com/index.html?datasheet=20444 This will bind a single epitope on a single protein. It may interfere somewhat with the rabbit antibody that follows it but chances are the rabbit antibody will recognize other proteins on the bacterium in addition to, or other than, what the monoclonal binds. Antibody ab20444 is also suitable for conjugation: it is purified and there are no other proteins in the solution. The order in which these antibodies are used is important. Do not apply the rabbit polyclonal ab20818 before the mouse monoclonal ab20444; the rabbit may very well hinder the mouse antibody. I hope this helps. Please let us know if you have any other questions or concerns.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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