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Our Abpromise guarantee covers the use of ab176106 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use a concentration of 1 µg/ml.
(or use at assay dependent concentration).
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 40, 42 kDa.|
Predicted MW (kDa): HuB/ELAVL2: 39.5 kDa, HuC/ELAVL3: 39.5kDa, HuD/ELAVL4: 41.7kDa
Why the observed band size is different than the predicted band size: HuC protein (ab91911) is a fragment and has an N terminal His tag and is predicted to present at 24 kDa.
Immunocytochemistry with SH-SY5Y cells stained with ab176106 in red and DAPI in blue, as a counter stain.
Fixation: Paraformaldehyde fixed (4%, 20 minutes).
Permeabilization: Triton X-100 permeabilized (0.1%, 10 minutes).
Blocking: 10% Goat Serum/PBS (1 hour).
Primary: ab176106, 1 µg/ml (10% Goat Serum/PBS, 2 hours at room temperature or over night at 4°C).
Secondary: Alexa 594 goat anti-mouse H+L used at a 1/1000 dilution in 1%Goat Serum/PBS for 1hour at RT.
Washing: 3X 1% Goat Serum/PBS, 10 minutes per wash.
DAPI: 2 uM in 1% Goat Serum/PBS, 10 minutes.
Specificity of antibody using the ELISA platform.
Antibody concentration is at 1µg/mL the concentration of recombinant proteins is at 250ng/mL. ab176106 reacts specifically to HuC/ELAVL3 and HuD/ELAVL4 proteins. Reactivity with HuB protein is predicted by the immunogen sequence similarity.
ab176106 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"