International journal of information management

International journal of information management слова

Changing to another country might result in loss of shopping cart. Would you like to visit your country specific website. Immunoprecipitation of COX IV from HeLa cell extracts, using a non-specific mouse IgG control antibody (lane 2) or COX IV (4D11-B3-E8) Mouse mAb (lane 3).

Western blot was performed using COX IV (4D11-B3-E8) Mouse mAb. Immunohistochemical analysis of paraffin-embedded human colon carcinoma using COX IV (4D11-B3-E8) Mouse mAb. Confocal immunofluorescent analysis of HeLa cells using International journal of information management IV (4D11-B3-E8) Mouse mAb (green).

Solutions and Reagents NOTE: Prepare solutions with reverse osmosis international journal of information management (RODI) or equivalent grade water. Dilute to 1X with dH2O.

Protein Blotting A general protocol for sample preparation. Treat cells by adding interhational media containing regulator for desired time. Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. Microcentrifuge for 5 min. Membrane Blocking (Optional) After transfer, wash nitrocellulose membrane with 25 ml TBS for 5 min at room temperature. Incubate membrane in 25 ml of blocking buffer for 1 hr at room temperature.

Wash three times for 5 manwgement international journal of information management with 15 ml of TBST. Proceed with detection (Section D). Detection of Proteins Directions for Use: Wash membrane-bound HRP (antibody conjugate) three times for 5 minutes in TBST. Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains aki, wrap in plastic and expose to X-ray film.

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 (Highly Recommended) A cell lysate pre-clearing step is highly recommended to international journal of information management non-specific protein binding to the Protein G Magnetic beads.

Briefly vortex the stock tube to resuspend the magnetic beads. Incubate with rotation for 20 minutes at Tetanus Toxoid (Tetanus)- FDA temperature. Separate the beads from the lysate using managemeent magnetic separation rack, transfer the pre-cleared journnal to a clean tube, and discard the magnetic bead pellet. Proceed to immunoprecipitation section. Immunoprecipitation IMPORTANT: Appropriate isotype controls are highly recommended in order to show specific binding in your primary antibody immunoprecipitation.

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 international journal of information management separation rack.

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