Overview

  • Product name
    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680)
    See all IgG secondary antibodies
  • Host species
    Donkey
  • Target species
    Rabbit
  • Specificity
    This antibody is specific to Rabbit IgG
  • Tested applications
    Suitable for: WBmore details
  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Conjugation
    Alexa Fluor® 680. Ex: 679nm, Em: 702nm

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark.
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • General notes

    We batch test Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680), ab175772 in fluorescent WB. Although we don’t batch test for ICC, ELISA, IHC-Fr or Flow cytometry customers have had success using Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680), ab175772 in these applications.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab175772 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 1/10000.

Images

  • All lanes : Anti-alpha Tubulin antibody (ab18251) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 5 µg
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 2 µg
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 1 µg

    Secondary
    All lanes : Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) (ab175772) at 1/10000 dilution

    Observed band size: 52 kDa
    why is the actual band size different from the predicted?



    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab18251 overnight at 4°C. Antibody binding was detected using ab175772 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • Cross-reactivity of the polyclonal secondary antibody ab182020 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. ab182020 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Goat anti-Donkey IgG H&L (HRP) (ab6988) was used at 1/20,000 dilution (50 µl/well), followed by incubation for 1h at RT.

    For the batch tested, ab182020 showed a cross-reactivity below 2% towards human IgG, mouse IgG, rat IgG, goat IgG and chicken IgY.

    This data was developed using the unconjugated antibody (ab182020).

References

This product has been referenced in:
  • Zhang M  et al. ß-Glucan from Saccharomyces cerevisiae induces SBD-1 production in ovine ruminal epithelial cells via the Dectin-1-Syk-NF-?B signaling pathway. Cell Signal 53:304-315 (2019). Read more (PubMed: 30401641) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

ICC/IF

Excellent
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
ab175772 - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) was used for immunological detection of motor protein dynein using Rabbit anti-dynein (ab75214) and ab175772 as secondary antibody (1:200 dilution- 1hr incubation @ RT).

Anu Shanu

Verified customer

Submitted Mar 27 2017

Abreviews
Application
Western blot
We used this secondary antibody for immunodetection of several antibodies on a LiCor Odyssee system and compared the signal with IRDye antibodies. Exemplary we show immunodetection of serine 6 phosphorylation of p53 (figure, part of this figure was used for the abreview of ab32132). The abcam ab175722 works as good as the IRDye antibody and give good and intense signals. We did not observe any unspecific binding to the membrane. (Dilution 1:10000 in 5% milk in TBS-T buffer, nitrocellulose membrane).

Mr. Christian Marx

Verified customer

Submitted Oct 28 2014

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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