Overview

  • Product name
    Anti-FITC antibody (Phycoerythrin)
    See all FITC primary antibodies
  • Description
    Sheep polyclonal to FITC (Phycoerythrin)
  • Host species
    Sheep
  • Conjugation
    Phycoerythrin. Ex: 488nm, Em: 575nm
  • Specificity
    Ab25539 recognises FITC.
  • Tested applications
    Suitable for: WB, ELISA, ICC/IF, Dot blotmore details
  • Immunogen

    Chemical/ Small Molecule: Fluorescein Isothiocyanate (Isomer I).

  • General notes


    Ab25539 shows cross absorption with mouse, human and rat serum proteins.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    Preservative: 0.09% Sodium azide
    Constituents: PBS, 16% Sucrose

    Also contains a stabilizing agent.
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    Ab25539 is purified by affinity chromatography.
  • Primary antibody notes
    Ab25539 shows cross absorption with mouse, human and rat serum proteins.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab25539 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
WB Use at an assay dependent dilution.
ELISA Use at an assay dependent dilution.
ICC/IF Use µg for 10 cells.
Dot blot Use at an assay dependent dilution.

Target

  • Relevance
    FITC (fluorescein isothiocyanate) is a fluorochrome dye that absorbs ultraviolet or blue light causing molecules to become excited and emit a visible yellow-green light. This emission ceases upon removal of the light causing the excitation. Fluorochrome labeling provides rapid, accurate localization of antigen-antibody interaction when one of the reactants is part of a cell, tissue or other biological structure. FITC is a commonly used marker for antibodies in immunofluorescent techniques since the conjugation of FITC to proteins is relatively easy and does not, in general, destroy the biological activity of the labeled protein. FITC is widely used as a hapten to label different proteins.
  • Alternative names
    • Fluorescein Isothiocyanate antibody

References

ab25539 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for contacting us. Yes, this antibody would be suitable for that purpose and has been tested for ICC/IF.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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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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