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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 use products containing Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or email@example.com.
Our Abpromise guarantee covers the use of ab150081 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.|
|ICC/IF||1/200 - 1/1000.|
|Flow Cyt||1/2000 - 1/4000.
ab199091-Rabbit monoclonal IgG (Alexa Fluor® 488), is suitable for use as an isotype control to complement this secondary antibody.
|IHC-P||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
ICC/IF image of ab8227 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab8227, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab150081 Alexa Fluor® 488 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.
The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.
Postnatal day 6 mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS. Sections were incubated with either (A) no primary antibody or (B ) anti-DDX4 (ab13840) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rabbit 488 (ab150081) applied at a 1/500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.
Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School
Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)
Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 µm thickness
Primary antibody 1: Rabbit anti cytokeratin 8 (ab53280), 1:100
Primary antibody 2: Rat anti-perlecan, 1:100
Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200
Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed, 1:200
Nuclei were counterstained with DAPI