Overview

  • Product name
    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 555)
    See all IgG secondary antibodies
  • Description
    Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 555)
  • Host species
    Donkey
  • Target species
    Rabbit
  • Specificity
    This antibody is specific to Rabbit IgG
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, ELISA, IHC-P, Flow Cytmore details
  • Immunogen

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

  • Conjugation
    Alexa Fluor® 555. Ex: 555nm, Em: 565nm

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
    The 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 ab150074 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.
ELISA Use at an assay dependent concentration. Use at an assay dependent dilution
IHC-P Use at an assay dependent concentration. Use at an assay dependent dilution
Flow Cyt 1/2000.

Images

  • ICC/IF image of ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to 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 ab150074 Alexa Fluor® 555 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.

    The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

  • 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:
  • Shi R  et al. Alteration of Adaptive Immunity in a Colorectal Peritoneal Carcinomatosis Model. J Cancer 10:367-377 (2019). Read more (PubMed: 30719130) »
  • Afroz S  et al. CGRP Induces Differential Regulation of Cytokines from Satellite Glial Cells in Trigeminal Ganglia and Orofacial Nociception. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30736422) »
See all 42 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer

Thank you for your enquiry.

Alexa Fluor 555 emits at 565 nm which is yellow-green.
The following table from our website may be useful:

https://www.abcam.com/ps/pdf/protocols/Fluorophore%20table.pdf

I hope this will be helpful. Please do not hesitate to let me know if you have any further questions.

Read More
Application
Immunocytochemistry/ Immunofluorescence
iPS derived neural progenitor cells were fixed in 4% formaldehyde (10 min) and then incubated in 1% serum, 0.1% triton, 0.1% BSA in PBS for 30 mins to permeabilise the cells and block non-specific binding. The cells were then incubated with Nestin antibody (ab105389, 1/200 dilution) for 1h at room temp. The secondary antibody (red) was Donkey Anti-Rabbit IgG H&L Alexa 555 (ab150074) at 1:500 dilution and incubated for 1h at room temp. Total cells were stained with DAPI (blue). Excellent positive Nestin staining was observed (red).

Abcam user community

Verified customer

Submitted Nov 28 2013

Application
Immunocytochemistry/ Immunofluorescence
Secondary antibody worked well at a 1:200 dilution. Clear staining observed when compared with staining done on cells not expressing human Nanog. Red colour did not fade under fluorescent light as I have seen with other red secondary antibodies previously. Primary antibody used here was Nanog (ab21624).

Dr. Sarah Ritson

Verified customer

Submitted Oct 07 2013

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