Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
Key features and details
- Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)
- Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm
- Host species: Donkey
- Isotype: IgG
- Suitable for: ICC/IF, Flow Cyt, IHC-P, ELISA, IHC-Fr
Overview
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Product name
Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488)
See all IgG secondary antibodies -
Description
Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) -
Host species
Donkey -
Target species
Rabbit -
Specificity
This antibody is specific to Rabbit IgG -
Tested applications
Suitable for: ICC/IF, Flow Cyt, IHC-P, ELISA, IHC-Frmore details -
Immunogen
The details of the immunogen for this antibody are not available.
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Conjugation
Alexa Fluor® 488. Ex: 495nm, Em: 519nm
Properties
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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: 23% Glycerol (glycerin, glycerine), PBS, 1% BSA -
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.
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Research areas
Images
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Immunocytochemistry/ Immunofluorescence - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
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 antibody (ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab150073 Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) used at 1µg/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.
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10min).
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Flow Cytometry - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)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® 488) (ab150073) was used at 1/4000 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 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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ELISA - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073) This image is courtesy of an Abreview submitted by Laura Comley.ab104603 staining DYNLL1 in mouse cervical spinal cord tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked with 10% serum for 1 hour at 21°C. Samples were incubated with primary antibody (2 µg/ml in PBS/10% serum/0.1% Triton X-100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG H&L (ab150073) (1/500) was used as the secondary antibody.
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Alexa Fluor® - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
