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40818 DATASHEET PDF

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Formaldehyde is toxic, use only in a fume hood. Blotting Membrane and Paper: Do not aliquot the antibody. Application Dilutions Western Blotting 1: Remove buffer once solution is clear. Scrape cells off the plate and transfer to microcentrifuge tubes. Ubiquitinating enzymes UBEs catalyze protein ubiquitination, a reversible process countered by deubiquitinating enzyme DUB action 1,2. The supernatant is the sample.

Incubate with rotation for 20 min at room temperature. To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold 1X PBS.

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Analyze cells in DNA staining solution on flow cytometer. Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently datashert only a limited amount of sample is available.

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Primary Antibody Dilution Buffer: Pre-wash magnetic beads just prior to use:. Volumes are for 10 cm x 10 cm cm 2 of membrane; for different sized membranes, adjust volumes accordingly.

Briefly vortex the stock tube to resuspend the magnetic beads. Sample Analysis Proceed to one of datashete following specific set of steps. This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity utilizing Protein A magnetic separation.

For best results, allow mountant to cure overnight at room temperature. Incubate 30 min on ice. Incubate specimen in fluorochrome-conjugated secondary antibody diluted in Antibody Dilution Eatasheet for 1—2 hr at room temperature in the dark.

Phospho-STING (Ser366) (D8K6H) Rabbit mAb #40818

Mizuno E et al. Resuspend cells in 4088. Proceed with detection Section D. 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.

To Purchase S View sizes. Solutions and Reagents Achieve higher quality immunofluorescent images using the efficient and cost-effective, pre-made reagents in our Immunofluorescence Application Solutions Kit NOTE: Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies: Incubate for 1 hr at room temperature.

Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. Protein A Magnetic Beads: Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature.

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More about how we get our images. To Purchase Dataasheet View sizes. Place the tube in a magnetic separation rack for seconds. Fix for 15 min at room temperature. Sonicate for 10—15 sec to complete cell lysis and shear DNA to reduce 4088 viscosity. Changing to another country might result in loss of shopping cart. The supernatant is the cell lysate. Treat cells by adding fresh media containing regulator for desired time. Transfer the lysate and antibody immunocomplex solution to the tube containing the pre-washed magnetic bead pellet.

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Immunoprecipitation for Native Proteins This datasheeg is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity utilizing Protein A magnetic separation. To prepare 10 ml, add 0. Immunoprecipitation Cell Lysate Pre-Clearing Optional A cell lysate pre-clearing step is highly recommended to reduce non-specific protein binding to the Protein A Magnetic beads. Incubate for at least 5 min at room temperature.

Microcentrifuge for 5 min. Prepare solutions with reverse osmosis deionized RODI or equivalently purified water.