Product nameDonkey Anti-Rabbit IgG H&L (Alexa Fluor® 405)
See all IgG secondary antibodies
SpecificityThis antibody is specific to Rabbit IgG
Tested applicationsSuitable for: IHC-Fr, Flow Cyt, IHC-P, ELISA, ICC/IFmore details
The details of the immunogen for this antibody are not available.
ConjugationAlexa Fluor® 405. Ex: 402nm, Em: 421nm
Storage instructionsShipped at 4°C. Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Please see notes section. Store In the Dark.
Storage bufferPreservative: 0.02% Sodium azide
Constituents: 23% Glycerol, PBS, 1% BSA
Concentration information loading...
PurityImmunogen affinity purified
We recommend storage time at 4°C should be minimal, since this may affect the signal strength.
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 firstname.lastname@example.org.
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150073)
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 555) (ab150074)
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150075)
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150076)
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 568) (ab175470)
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 750) (ab175731)
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) (ab175772)
- Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175780)
- Donkey Anti-Rabbit IgG H&L (ab182020)
- Donkey Anti-Rabbit IgG H&L (HRP) (ab205722)
- Donkey Anti-Rabbit IgG H&L (Biotin) (ab207999)
Our Abpromise guarantee covers the use of ab175651 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|ICC/IF||1/200 - 1/1000.
We recommend the use of a dedicated 405 filter for optimal results not the DAPI filter. The DAPI filter may not excite until the maximum emission peaks of Alexa Fluor® 405 dye (see difference below)
Section of adult murine femur was stained with rabbit anti-fibronectin (ab23750) and detected with donkey anti-rabbit AlexaFluor405 (ab175651) - shown in green. Counterstain shows CD31+ blood vessels and cells (red) and Sca1+ arterioles and cells (blue).
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, blocked with Image-iT® FX Signal Enhancer at room temperature for 30 min 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 (blue) was ab175651 Alexa Fluor® 405 donkey anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DRAQ5 (ab108410) was used to stain the cell nuclei (red) at a concentration of 1.25µM.
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).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab175651 has been referenced in 13 publications.
- Brasileiro AD et al. Effects of diabetes mellitus on myenteric neuronal density and sodium channel expression in the rat ileum. Brain Res 1708:1-9 (2019). PubMed: 30500400
- Kimble JB et al. Antibody-Mediated Protective Mechanisms Induced by a Trivalent Parainfluenza Virus-Vectored Ebolavirus Vaccine. J Virol 93:N/A (2019). PubMed: 30518655
- Suzuki IK et al. Human-Specific NOTCH2NL Genes Expand Cortical Neurogenesis through Delta/Notch Regulation. Cell 173:1370-1384.e16 (2018). PubMed: 29856955
- Mayeuf-Louchart A et al. MuscleJ: a high-content analysis method to study skeletal muscle with a new Fiji tool. Skelet Muscle 8:25 (2018). PubMed: 30081940
- Niquille M et al. Neurogliaform cortical interneurons derive from cells in the preoptic area. Elife 7:N/A (2018). PubMed: 29557780