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Find answers on our FAQs page. Would you like to visit your country specific website? Detection of Proteins Directions for Use: Each of the fragile X proteins can self-associate, as well as form heteromers with the other two related proteins 3. Application Dilutions Western Blotting 1: Biotinylated Protein Ladder Detection Pack: Aspirate media from cultures; wash cells with 1X PBS; 51161.
Microcentrifuge for 5 min.
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Primary Antibody Dilution Buffer: Incubate datashret with membrane for 1 minute, remove excess solution membrane remains wetwrap in plastic and expose to X-ray film.
FXR2 Antibody – More about how we get our images. Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube.
Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature. From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: Volumes are for 10 cm x 10 cm cm 2 of membrane; for different sized membranes, adjust volumes accordingly. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody. Fragile X syndrome is a genetic disorder characterized by a spectrum of physical and behavioral features and is a frequent form of inherited mental retardation 1.
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended.
Electrotransfer to nitrocellulose membrane Wash three times for 5 min each with 15 ml of TBST.
To Purchase S View sizes. These results suggest that fragile X syndrome is related to abnormal translation caused by defects in RNAi-related pathways.
Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human FXR2 protein.
Protein Blotting A general protocol for sample preparation. Blotting Membrane and Paper: Do not aliquot the antibody.
Please refer to primary antibody datasheet or product webpage for recommended antibody dilution. Sonicate for 10—15 sec to complete cell lysis and shear DNA to reduce sample viscosity.
Dilute to 1X with dH 2 O. datasgeet
Solutions and Reagents From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: Proceed with detection Section D. Antibodies are purified by protein A and peptide affinity chromatography.
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Treat cells by adding fresh media containing regulator for desired time. Prepare solutions with reverse osmosis deionized RODI or equivalently purified water.
Changing to another country might result in loss of shopping cart. Additionally, it is recommended that you verify the adtasheet of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. Prepare solutions with reverse osmosis deionized RODI or equivalent grade water.