Overview

  • Product name
    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 568)
    See all IgG secondary antibodies
  • Host species
    Donkey
  • Target species
    Rabbit
  • Specificity
    This antibody is specific to Rabbit IgG
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, IHC-P, Flow Cyt, ELISAmore details
  • Immunogen

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

  • Conjugation
    Alexa Fluor® 568. Ex: 578nm, Em: 603nm

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.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituents: 30% Glycerol, 1% BSA, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    This antibody was isolated by affinity chromatography using antigen coupled to agarose beads.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • General notes

    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 ab175470 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 Use at an assay dependent concentration.
ICC/IF 1/200 - 1/1000.
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/2000.
ELISA Use at an assay dependent concentration.

Images

  • ICC/IF image of ab6046 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1% BSA/10% normal donkey serum/0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (yellow) was ab175470, Alexa Fluor® 568 Donkey anti-Rabbit IgG - H&L, used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    Alexa Fluor® 568 is pseudocolored in yellow.

  • HeLa cells showing negative staining by ICC/IF using only secondary antibody. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The secondary antibody (yellow) was ab175470, Alexa Fluor® 568 Donkey anti-Rabbit IgG - H&L, used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

     

    Alexa Fluor® 568 is pseudocolored in yellow.

  • 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:
  • Huang X  et al. Neuronal complement cascade drives bone cancer pain via C3R mediated microglial activation. Brain Res 1698:81-88 (2018). Read more (PubMed: 29909203) »
  • Koukouli F  et al. Nicotine reverses hypofrontality in animal models of addiction and schizophrenia. Nat Med 23:347-354 (2017). Read more (PubMed: 28112735) »
See all 6 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
This secondary ab works as expected. I used the dilution 1in1000.
The picture shows a picture where mouse cells (engineered to express mCherry) were stained with the primary antibody for mCherry (ab213511) and subsequently stained with ab175470.

The picture was taken with a Leica Microscope using a 20X objective.

Dr. Francesco Elia Marino

Verified customer

Submitted Sep 26 2018

Application
Flow Cytometry
Immediately after cell isolation I stained all cells with ab175470, as expected it stained all smooth muscle cells in the sample (13%) ab53219 was used as primary antibody. On the histogram can be seen the fraction of positive stained smooth muscle cells. The antibody worked perfectly fine.

Abcam user community

Verified customer

Submitted Jul 10 2017

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