Blocking Buffer 37C Phase

On Friday (April 11th), I ran the second phase of my blocking buffer experiment! This consisted of blocking each solution at an incubation temperature of 37˚C. Due to a lack of cut slides, I slightly modified the solutions for this run. For this phase, I used 1/2x and 1x casein at 30 minute and 1 hour blocking times and 0.2%, 2.5%, and 5% BSA at 30 minute and 1 hour blocking times. I gathered all of my materials and went to the 37˚C room down the hall.

 For this phase, I used the same protocol from the room temperature phase I ran on February 28th, as seen below.

1) Add 1mL of the specified protein solution to the specified slide.
2) Block on the shaker for the specified time.
3) Drain well with pipette into waste container.
4) Wash slide with 4 mL PBS for 5 minutes.
5) Drain well with pipette into waste container.
6) Transfer slide to new plate.
7) Stain with 4 mL blue stain for 15 minutes.
8) Drain stain into waste.
9) Wash slide with 4 mL PBS for 5 minutes.
10) Air dry slides for 5 minutes on paper towel.
11) Write blocking solution, blocking time, and temperature on back of slides.

 I will be away on college trips next week, so I will not be able to go to the lab Friday April 28th. I look forward to imaging the slides from this phase when I get back to the lab on April 25th!