Overview

  • Product name

    Anti-Protein L antibody (FITC)
    See all Protein L primary antibodies
  • Description

    Chicken polyclonal to Protein L (FITC)
  • Host species

    Chicken
  • Conjugation

    FITC. Ex: 493nm, Em: 528nm
  • Tested applications

    Suitable for: IHC-Frmore details
  • Species reactivity

    Reacts with bacteria.
  • Immunogen

    Purified recombinant protein L.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab63504 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
IHC-Fr 1/500 - 1/1000.

Target

  • Relevance

    Protein L is a 36,000 dalton immunoglobulin-binding protein isolated from the bacteria Peptostreptococcus magnus. Unlike Protein A and Protein G, which bind to the Fc region of immunoglobulins (antibodies), Protein L binds antibodies through light chain interactions. Since no part of the heavy chain is involved in the binding interaction, Protein L binds a wider range of antibody classes than Protein A or G. Protein L binds to representatives of all antibody classes, including IgG, IgM, IgA, IgE and IgD. Single chain variable fragments (ScFv) and Fab fragments also bind to Protein L. Despite this wide binding range, Protein L is not a universal antibody-binding protein. Protein L binding is restricted to those antibodies that contain kappa light chains and it is only effective in binding certain subtypes of kappa light chains - about 65% of human immunoglobulins carry kappa light chains. Given these specific requirements for effective binding, the main application for immobilized Protein L is purification of monoclonal antibodies from ascites or cell culture supernatant that are known to have the kappa light chain. Protein L is extremely useful for purification of VLkappa-containing monoclonal antibodies from culture supernatant because it does not bind bovine immunoglobulins, which are often present in the media as a serum supplement. Also, Protein L does not interfere with the antigen-binding site of the antibody, making it useful for immunoprecipitation assays, even using IgM.
  • Cellular localization

    Cell surface
  • Alternative names

    • ProteinL antibody
    • RPL antibody

References

ab63504 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 your enquiry.

Currently, we have 4 antibodies against Protein L but none of them have been tested in immunoprecipitation:

ab63503, ab63504, ab63505, ab63506

If your customer would like to test any of these antibodies in IP and get a Discount code from us, please do let me know. The Terms and Conditions of this testing offer can be found at: https://www.abcam.com/collaborationdiscount.

If you need anything further or any help then please let me know.

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Answer

Thank you for contacting us. Unfortunately, I would not recommend using a chicken antibody against a different target (such as ab1394 or ab63504), as it cannot be guaranteed that they do not react with any human antigen. I would rather suggest to use a chicken IgY polyclonal isotype control, as these isotype controls are specifically tested not to cross react. As such we currently have ab37382 available (Click here (or use the following: https://www.abcam.com/index.html?datasheet=37382).). As this antibody however is not labelled with FITC, I may recommend our EasyLink Conjugation kit (ab102883 - ab102885, see below and attached). For the compensation and set-up of the flow cytometer, I would suggest to use ab63498 to ensure the optimal settings. EasyLink antibody conjugation kits: https://www.abcam.com/easylink Click here (or use the following: https://www.abcam.com/index.html?datasheet=102883). Having said this however, I may let you know that isotype controls do not really represent a good negative control. The reasons become apparent in your particular case: The control antibody is of a different preparation ('chicken', or in case of monoclonals a different clone) and therefore may show varying binding properties compared to the specific antibody of interest (ab63498). Furthermore, as the conjugation conditions may have been different, the labelling with the fluorochrome can vary which can be a very important issue in flow cytometry. As a result of different antibody-to-fluorochrome ratios and/or different fluorochrome batches, the control antibody might indeed show a very different staining pattern, and even if it does not bind at all (as it is supposed to) can for these reasons not directly be compared to the antibody of interest. Instead, in flow cytometry another type of control is often used to determine positivity for the marker of interest. This control is termed fluorescence minus one or FMO and works like an isotype control without the actual antibody. FMO controls are set up by leaving out one of the antibodies in your staining panel. You may find more information about this type of control in the latest publications or on flow cytometry websites, such as http://www.flowcytometryuk.org/, http://www.cyto.purdue.edu/ or http://www.isac-net.org . Plus, your local flow cytometry community (core facility?) might be happy to help and advise in the use of optimal controls which are required for publishing the results. 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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