S TRANSISTOR (PNP). FEATURE. Excellent hFE linearity. MAXIMUM RATINGS (TA=25℃ unless otherwise noted). Symbol. Parameter. Value. Units. Order Number Normal Lead Free Plating S-x-TK SL-x-TK Package SOT TO Pin Details, datasheet, quote on part number: S. Part, S-TO Category. Description, LOW Voltage HIGH Current Small Signal PNP Transistor. Company, Unisonic Technologies. Datasheet, Download .
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Please refer to primary antibody datasheet or dataxheet 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 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.
MKK6 (D31D1) Rabbit mAb #8550
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 attributed 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. This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity utilizing Protein A magnetic separation. A cell lysate pre-clearing step is highly recommended to reduce non-specific protein binding to the Protein A Magnetic beads. Pre-clear enough lysate for test samples and isotype controls. Pre-wash magnetic beads just prior to use:.
Carefully remove the buffer once the solution is clear.
datashest Place tube back in magnetic separation rack. Remove buffer once solution is clear. Repeat washing step once more. The optimal lysate concentration will depend on the expression level of the protein of interest.
Appropriate isotype controls are highly recommended in order to show specific binding in your primary antibody immunoprecipitation. Isotype controls should be concentration matched and run alongside the primary antibody samples. Do not aliquot the antibody.
Monoclonal antibody is produced by immunizing animals with recombinant, full-length MKK6 expressed in E. MKK3 and MKK6 are both activated by different forms of cellular stress and inflammatory cytokines 4,5. Changing to another country might result in loss of shopping cart. Would you like to visit your country specific website?
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 water.
Dilute to 1X with dH 2 O. Primary Antibody Dilution Buffer: Biotinylated Protein Ladder Detection Pack: Blotting Membrane and Paper: Protein Blotting A general protocol for sample preparation.
s datasheet pdf
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 8550s 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 min each with 15 ml of TBST.
S Datasheet, Equivalent, Cross Reference Search. Transistor Catalog
Proceed with detection Section D. Detection of Proteins Directions datashet Use: Incubate substrate with membrane for 1 minute, remove excess solution membrane remains wetwrap in plastic and 8550s to X-ray film. Immunoprecipitation for Native Proteins This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity utilizing Protein A magnetic separation.
Protein A Magnetic Beads: ATP 10 mM for kinase assays: Preparing Cell Lysates Aspirate media.
To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold 1X PBS. Remove PBS and add 0. Scrape cells off the plate and transfer to microcentrifuge tubes. Sonicate on ice three times for 5 sec each. The supernatant is the cell lysate. Immunoprecipitation Cell Lysate Pre-Clearing Optional A cell lysate pre-clearing step datashwet highly recommended to reduce non-specific protein binding to the Protein A Magnetic beads.
Briefly vortex the stock tube 8550w resuspend the magnetic beads. Pre-wash magnetic beads just prior to use: Place the tube in a magnetic separation rack for seconds. Incubate with rotation for 20 minutes at room temperature. Separate the beads from the lysate using a magnetic separation rack, transfer the pre-cleared lysate to a clean tube, and discard the magnetic bead pellet.
Proceed to immunoprecipitation section. Pre-wash magnetic beads see Cell Lysate Pre-Clearing section, steps 1 and 2. Transfer the lysate and antibody immunocomplex solution to the tube containing the pre-washed magnetic bead pellet. Incubate with rotation for 20 min at room temperature.
Pellet beads using magnetic separation rack. Keep on ice between washes.
Proceed to analyze by western immunoblotting or kinase activity section D. Sample Analysis Proceed to one of the following specific set of steps.
Transfer the supernatant to a new tube. The supernatant is the sample. Analyze sample by western blot see Western Immunoblotting Protocol. Vortex, then microcentrifuge for 30 sec.
Transfer supernatant containing phosphorylated substrate to another tube. Application Dilutions Western Blotting 1: Human, Mouse, Rat, Monkey. To Purchase S View sizes. Find datasheey on our FAQs page.