...PBS on a rocking platform. Permeabilize cells for 10 min at room temperature ( RT ) on a rocking platform...PBS on a rocking platform. Section 3: Labeling with antibody Block for 20 min at RT on a rocking platform...platform in 500 µL blocking buffer. Remove the blocking buffer and dispose of it in an appropriate waste container...Equipment Pipette controller Pipette tips and pipettes Rocking platform Tweezers Fluorescent microscope 0.5–10...buffer: Dilute 20 µL of Triton X-100 in 10 mL PBS. Blocking buffer: Dilute 0.5 g BSA and 30 µL Triton X-100...container. Wash 3x for 5 min in 500 µL PBS on a rocking platform. Dilute the fluorescently-labeled secondary...container. Wash 3x for 5 min in 500 µL PBS on a rocking platform. (Optional) Counterstain nuclei with 500...

...information specific to your antibody, such as ideal blocking buffer and optimal antibody concentrations. Consider...When the run is complete, select Done. Section 5: Blocking Prepare 1X TBST as follows: 25 mL of 20X TBS 2.5...20 472.5 mL of deionized water Mix well Prepare blocking buffer as follows: Dilute 5% w/v non-fat milk ...milk into 100 mL of 1X TBST. Pro-Tip The ideal blocking buffer will vary between antibodies. Refer to ...1X TBST, protein side up. Block the membrane in blocking buffer for 1 h at RT on a shaking platform. Wash...primary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal concentration will...membrane overnight in primary antibody at 4 °C on a rocking platform. Pro-Tip Primary antibody incubation can...

...Microcentrifuge Electrophoresis chamber Power supply Rocking platform Fume hood Metal spatula Razor blade Plastic...deionized water for 5 min with gentle agitation on a rocking platform. Pour off the water in the sink. Add 20...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the SimplyBlue SafeStain in ...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Add 100...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Take ...

...Workflow Timeline Day 1: Antigen Coating Day 2: Blocking Day 3: Primary antibody incubation Day 4: Secondary...aspirate the wash buffer from the wells. Prepare the blocking buffer (1% BSA in PBS) as follows: Add 250 mg ...mix. Using a multichannel pipette, add 200 µL of blocking buffer to each well. Cover the plate with a plate...

...DNA. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 15–...DMEM complete. Incubate the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X...

...it has been diluted. Mix by swirling or gently flicking the tube. Incubate for 5 minutes at room temperature...the walls of the tube. Mix by swirling or gently flicking the tube. f. Incubate for 20-30 minutes at room...

.... You can find Addgene's sequencing results by clicking on the "View Sequences" link on the Plasmid Information...

... microcentrifuge or falcon tube. GENTLY mix by flicking the bottom of the tube with your finger a few ...

...ligase, 5min at room temperature is enough. For trickier ligations (such as ligation of annealed oligos...

...mixture. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 12–...

...dilutions. Pro-Tip Make sure that the qPCR is valid by checking to the following: Standard curve: R 2 (coefficient...