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Solutions and Reagents From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: Would you like to visit your country specific website? This can be done by re-exposing the blot to ECL reagents and making sure dataasheet is no signal prior to adding the next primary antibody. Volumes are for 10 cm x 10 cm cm 2 of membrane; for different sized membranes, adjust volumes accordingly.
Please refer to primary antibody datasheet or product webpage for recommended antibody dilution. Primary Antibody Dilution Buffer: Datasjeet Antibody – Application Dilutions Western Blotting 1: Prepare solutions with reverse osmosis deionized RODI or equivalent grade water.
Wash three times for 5 min each with 15 ml of TBST. Electrotransfer to nitrocellulose membrane Find answers on our FAQs page. Treat cells by adding fresh media containing regulator for desired time.
Polyclonal Antibody – FXR2 Antibody – Western Blotting, UniProt ID P51116, Entrez ID 9513 #4247
Detection of Proteins Directions for Use: It should be noted that for the best possible results a fresh blot is always recommended. Incubate substrate with membrane for 1 minute, remove excess solution membrane remains wetwrap in plastic and expose to X-ray film.
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. Prepare solutions with reverse osmosis deionized RODI or equivalently purified water.
Each of the fragile X proteins can self-associate, as well as form heteromers with the other two related proteins 3. Proceed with detection Section D. Protein Blotting A general protocol for sample preparation.
Aspirate media from cultures; wash cells with 1X PBS; aspirate. To Purchase S View sizes. These results suggest that fragile X syndrome is related to abnormal translation caused by defects in RNAi-related pathways.
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Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature. Changing to another country might result in loss of shopping cart.
Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. Additionally, it is recommended that you verify the removal 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.
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Sonicate for 10—15 sec to complete cell lysis and shear DNA to reduce sample viscosity. Antibodies are purified by protein A and peptide affinity chromatography. Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available.
Biotinylated Protein Ladder Detection Pack: Dilute to 1X with dH 2 O. Do not aliquot the antibody. From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human FXR2 protein.
Blotting Membrane and Paper: Microcentrifuge for 5 min.