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Western blot analysis of extracts from various cell types using Flotillin-2 L Antibody.
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Please refer to primary antibody datasheet or product webpage for recommended antibody dilution. 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. Reprobing of an existing membrane is a convenient means to immunoblot for datsheet 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.
Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal l2294 to binding of the new antibody is not leftover signal from the first immunoblotting experiment.
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. Prepare solutions with reverse osmosis deionized RODI or equivalently purified water. Do not aliquot the antibody.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding residue surrounding Leu of human flotillin Antibodies are purified by peptide affinity chromatography. Flotillins belong to a family of lipid raft-associated integral membrane proteins that carry an evolutionarily conserved domain called the prohibitin homology domain PHB 1.
Flotillin members are ubiquitously expressed and located in noncaveolar microdomains lipid rafts on the plasma membrane where they support signal transduction and regulate lipid raft motility and localization Two flotillin members have been described, flotillin-1 and flotillin Flotillin-2 is mainly localized to the plasma membrane and is prevalent in cell-cell contact sites.
However, overexpressed flotillin-2 has also been found in the late endosome 4,8,9. Both flotillin-1 and flotillin-2 are commonly used as lipid raft-associated markers.
Datasheeet to another country might result in loss of shopping cart. Would you like to visit your country specific website? Flotillin-2 L Antibody – More about how we get our images. Solutions and Reagents From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: Prepare solutions with reverse osmosis deionized RODI or equivalent grade datasheet.
Dilute to 1X with dH 2 O. Dataheet Antibody Dilution Buffer: Biotinylated Protein Ladder Detection Pack: Blotting Membrane and Paper: Protein Blotting A general protocol for sample preparation.
Treat cells by adding fresh media containing regulator for desired time. Aspirate media from cultures; wash cells with 1X PBS; aspirate. Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube.
Sonicate for 10—15 sec to complete cell lysis and shear DNA to reduce sample viscosity.
Microcentrifuge for 5 min. Electrotransfer to nitrocellulose membrane Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature. Wash three times for 5 darasheet each with 15 ml of TBST. Proceed with detection Section D.
L 데이터시트(PDF) – STMicroelectronics
Detection of Proteins Directions for Use: Incubate substrate with membrane for 1 minute, remove excess solution membrane remains wetsatasheet in plastic and expose to X-ray film. Application Dilutions Western Blotting 1: Flotillin-2 L Antibody detects endogenous levels of total flotillin-2 protein. Human, Mouse, Rat, Monkey. To Purchase S View sizes. Find answers on our FAQs page.