Overview

  • Product name
    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594)
    See all IgG secondary antibodies
  • Description
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594)
  • Host species
    Goat
  • 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

    Other Immunogen Type corresponding to Rabbit IgG.

  • Conjugation
    Alexa Fluor® 594. Ex: 590nm, Em: 617nm

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 ab150080 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 - 1/4000.

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 donkey 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 primary antibody (ab6046, 5µg/ml) overnight at +4°C. The secondary antibody (orange) was ab150080 Alexa Fluor® 4594 goat anti-rabbir 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.

  • Overlay histogram showing Jurkat cells stained with ab40763 (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 goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40763, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 594) (ab150080) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (ab172730, 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 561nm laser and 610/20 bandpass filter.

  • Cross-reactivity of the polyclonal secondary antibody ab182016 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. ab182016 was then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT.

    For the batch tested, ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, Rat IgG and Chicken IgY.

    This data was developed using the unconjugated antibody (ab182016).

  • Cross-reactivity of Goat anti-Rabbit IgG H&L (ab182016) and Goat anti-Rabbit IgG H&L obtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 µg/ml (50 µl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 µg/ml and gradually diluted 1/4 (50 µl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (ab6885) was used at 1/10,000 dilution (50 µl/well), followed by incubation for 1h at RT. This data is from a representative dilution.

    This data was developed using the unconjugated antibody (ab182016).

  • IHC-P image of alpha smooth muscle actin (ab5694) staining E16.5 mouse embryo gut. Paraformaldehyde fixed and paraffin embedded E16.5 mouse embryo gut sections were dewaxed and rehydrated before antigen retrieval (4 mins in a pressure cooker in 10mM Tris/0.4mM EDTA buffer pH 9.5). They were then incubated in 50mM NH4Cl for 30 minutes and washed/blocked in 3x 10 minute washes of PBS containing 1% BSA + 0.2% gelatine and 0.05% saponin. Sections were incubated overnight with a primary antibody against alpha smooth muscle actin (ab5694), diluted 1/250 in PBS containing 0.1% BSA and 0.3% triton. After 3 x 10 minute washes in of PBS containing 0.1% BSA, 0.2% gelatine and 0.05% saponin, the sections were incubated for 1 hr in the secondary antibody (ab150080, diluted 1/400, shown in red) and then the 3 washes repeated. Sections were mounted in Vectashield with DAPI (blue).

     

References

This product has been referenced in:
  • Wu M  et al. Synergistic effects of sulfur dioxide and polycyclic aromatic hydrocarbons on pulmonary pro-fibrosis via mir-30c-1-3p/ transforming growth factor ß type II receptor axis. Chemosphere 219:268-276 (2019). Read more (PubMed: 30543962) »
  • Kong F  et al. 5-Hydroxymethylfurfural Mitigates Lipopolysaccharide-Stimulated Inflammation via Suppression of MAPK, NF-?B and mTOR Activation in RAW 264.7 Cells. Molecules 24:N/A (2019). Read more (PubMed: 30642099) »
See all 53 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Paraformaldehyde fixed and paraffin embedded E16.5 mouse embryo gut sections were dewaxed and rehydrated before antigen retrieval (4 mins in a pressure cooker in 10mM Tris/0.4mM EDTA buffer pH=9.5). They were then incubated in 50mM NH4Cl for 30 minutes and washed/blocked in 3x 10 minute washes of PBS containing 1% BSA + 0.2% gelatine and 0.05% saponin. Sections were incubated overnight in primary antibody against alpha smooth muscle actin (ab5694), diluted 1/250 in PBS containing 0.1% BSA and 0.3% triton. After 3 x 10 minute washes in of PBS containing 0.1% BSA, 0.2% gelatine and 0.05% saponin, the sections were incubated for 1 hr in the secondary antibody (ab150080, diluted 1/400) and then the 3 washes repeated. Sections were mounted in Vectashield with DAPI.
Images are Alexa594, Dapi and the merge.

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Submitted Nov 04 2013

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