Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150063)

Overview

  • Product name

    Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed
    See all IgG secondary antibodies
  • Description

    Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 647), pre-adsorbed
  • Host species

    Donkey
  • Target species

    Rabbit
  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with rabbit IgG and with light chains common to other rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, goat, human, mouse, pig and rat IgG was detected. This antibody may cross react with IgG from other species.

     

  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cyt, ELISA, IHC-Frmore details
  • Minimal
    cross-reactivity


    Human, Mouse, Rat more details
  • Immunogen

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

  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm

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. Stable for 12 months at -20°C. Store In the Dark.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: 1% BSA, 30% Glycerol, PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Antiserum was cross adsorbed using a human, mouse and rat immunosorbents to remove cross reactive antibodies. The antibody to rabbit IgG 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

Associated products

Applications

Our Abpromise guarantee covers the use of ab150063 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
ICC/IF 1/200 - 1/1000.
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/1000 - 1/2000.

ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control to complement this secondary antibody.

 

ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Images

  • Overlay histogram showing Jurkat cells stained with ab16669 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal donkey serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Donkey anti-rabbit IgG H&L (Alexa Fluor® 647) (ab150063) was used at 1/1000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635nm) and 675/30 bandpass filter.

  • ICC/IF image of ab6046 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat 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 (red) was ab150063 Alexa Fluor® 647 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.
  • 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 goat 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 (red) was ab150063 Alexa Fluor® 647 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.

References

This product has been referenced in:

  • Hiraga SI  et al. Human RIF1 and protein phosphatase 1 stimulate DNA replication origin licensing but suppress origin activation. EMBO Rep 18:403-419 (2017). Flow Cyt . Read more (PubMed: 28077461) »
  • Xu X  et al. MicroRNA-1906, a Novel Regulator of Toll-Like Receptor 4, Ameliorates Ischemic Injury after Experimental Stroke in Mice. J Neurosci 37:10498-10515 (2017). Read more (PubMed: 28924010) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abreviews
Application
Immunohistochemistry (PFA fixed)
We have used this antibody together with the antibody cytokeratin 14 (ab51054) on skin sections.
The working concentration was 1/100 for 30 minutes at room temperature.

Abcam user community

Verified customer

Submitted Feb 14 2018

Answer

I can recommend the following APC conjugated product (https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-hl-allophycocyanin-pre-adsorbed-ab130805.html) for use with ab134118. I have also included two of our Alexa 647 antibodies (https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-hl-alexa-fluor-647-pre-adsorbed-ab150083.html & https://www.abcam.com/donkey-polyclonal-secondary-antibody-to-rabbit-igg-hl-alexa-fluor-647-pre-adsorbed-ab150063.html) which have emmision/excitation in the same range as APC. I have included these because they have lower cost, greater size and have been pre absorbed against rat, mouse and human. All three products are guaranteed under our AbPromise for use in flow. Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Allophycocyanin), pre-adsorbed (ab130805) https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-hl-allophycocyanin-pre-adsorbed-ab130805.html Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 647), pre-adsorbed https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-hl-alexa-fluor-647-pre-adsorbed-ab150083.html Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 647), pre-adsorbed https://www.abcam.com/donkey-polyclonal-secondary-antibody-to-rabbit-igg-hl-alexa-fluor-647-pre-adsorbed-ab150063.html Abcam also carries other products essential for Flow Cytometry such viability controls like 7-AAD or Propidium Iodide. Propidium Iodide (PI) binds to double-stranded DNA. PI cannot cross intact plasma membrane and therefore will only be present in DNA of cells where the plasma membrane has been compromised/ permeabilized. We do carry PI and offer a 7-AAD Cell-Mediated Cytotoxicity Assay Kit. Propidium Iodide: https://www.abcam.com/Propidium-Iodide-ab14083.html 7-AAD Kit: https://www.abcam.com/7-AADCFSE-Cytotoxicity-Assay-Kit-ab133073.html

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