Goat anti-rabbit IgG H&L (Alexa Fluor® 555) is a secondary antibody with a maximum absorption wavelength of 555nm and a maximum emission wavelength of 565nm. Ideal for fluorescent cell and tissue imaging. Suitable for ICC/IF, IHC, flow cytometry and ELISA.
- Intense fluorescence and high photostability allowing more time for image capture especially when detecting low abundance targets
- Minimal spectral overlap with other Alexa Fluor® dyes makes this ideal for use in multi-color analysis
- Cited in over 260 publications
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application IHC-Fr | Reactivity Reacts | Dilution info - | Notes - |
Application Flow Cyt | Reactivity Reacts | Dilution info 1/2000 | Notes - |
Application ICC/IF | Reactivity Reacts | Dilution info 1/200.00000 - 1/1000.00000 | Notes - |
Application IHC-P | Reactivity Reacts | Dilution info - | Notes Use at an assay dependent dilution |
Application ELISA | Reactivity Reacts | Dilution info - | Notes Use at an assay dependent dilution |
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Ig gamma 1 chain C region, Ig gamma 3 chain C region, Ig gamma 4 chain C region, Ig gamma chain C region, Ig gamma-2 chain C region, Ig kappa-b4 chain C region, Ig kappa-b5 chain C region, Ig kappa-b9 chain C region, Ig lambda chain C region, Immunoglobin heavy constant gamma 1, Immunoglobulin G, K-BAS
Goat anti-rabbit IgG H&L (Alexa Fluor® 555) is a secondary antibody with a maximum absorption wavelength of 555nm and a maximum emission wavelength of 565nm. Ideal for fluorescent cell and tissue imaging. Suitable for ICC/IF, IHC, flow cytometry and ELISA.
- Intense fluorescence and high photostability allowing more time for image capture especially when detecting low abundance targets
- Minimal spectral overlap with other Alexa Fluor® dyes makes this ideal for use in multi-color analysis
- Cited in over 260 publications
Preservative: 0.02% Sodium azide
Constituents: PBS, 23% Glycerol (glycerin, glycerine), 1% BSA
The antibody was isolated by affinity chromatography using antigen coupled to agarose beads.
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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.
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This data was done using a primary antibody to Calmegin (EPR11832) Anti-Calmegin antibody [EPR11832] ab171971. The primary antibody concentration was 1: 100 and the secondary antibody concentration was 1:500.
ICC/IF image of Anti-beta Tubulin antibody - Loading Control 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 (Anti-beta Tubulin antibody - Loading Control ab6046, 1μg/ml) overnight at +4°C. The secondary antibody (yellow) was ab150078 Alexa Fluor® 555 goat 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 (yellow) was ab150078 Alexa Fluor® 555 goat 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.
Cross-reactivity of the polyclonal secondary antibody Goat Anti-Rabbit IgG H&L 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. Goat Anti-Rabbit IgG H&L 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) (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, Goat Anti-Rabbit IgG H&L 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 (Goat Anti-Rabbit IgG H&L ab182016).
Cross-reactivity of Goat anti-Rabbit IgG H&L (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) (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 (Goat Anti-Rabbit IgG H&L ab182016).
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