And it just keeps piling on...

It's amazing how many posts I am now doing now that I know that at least one person is reading this... I don't expect this to last though as I think it's only because I'm not really pleased with things yet. I just ran another experiment, and found that my B cells are not actually expressing CD4, which is good, but I spoke to the company rep and he was literally shocked at the theory as to why I think that was happening... OK, it wasn't my theory, but the theory from the girl next door, and she was right... I just tested it, and she's completely right. Bloody sticky B cells... Not happy...

Now let's see the ramifications of this....

First, I spent months working out the method for this separation step. Which you can see below... On the left 2 different cell types in one solution, same with the middle, but the third, is the cells that I pulled out of the mixture using the aforementioned antibody... Beautifully separated cells, but the wrong ones...

I worked more months getting the gene array working properly. I spent thousands of dollars running the experiments. And then I spent countless hours trying to figure out how to analyze it (the guy who was paid to do the analysis by administration warned them months in advance that he was leaving last October, and here it is May and they still don't have anyone). All to find out that it doesn't look like I can use it....

Good points from this observation: I learned that mouse B-cells need to be blocked with a solution of 1% of each, normal rat, rabbit, serum (with 1% foetal calf serum thrown in for good luck), and 10uL per 5mL of 2.4g2 antibody to help block the B-cells Fc receptors. I also learned that the anti-CD4 antibody (L3T4 clone) that I was using sticks to B-cells in every way possible besides the classical way antibodies are supposed to bind, ie it will bind to B-cell on its side, and upside down, but not likely right side up. Another good point, I have to re-do it anyway to give some sort of story to this chapter in my thesis. The experiments I did last year showed that there was no change, which admittedly raised an eyebrow, but it actually ends up being a good result because it suggests that our T cell drug is not doing anything to the antigen presenting B cell. I just can't get that published anywhere because no one will publish something based on a purification of B-cells using a CD4 antibody... Sorry, that's another crap thing, wrong paragraph.Well, I may do another post tomorrow because I am writing from home. Oh ya though, I bought a lottery ticket today. The prize is 22million. I have won the last 4 out of 5 times I have played, (only between $20-$40 or $50) but today I am feeling pretty crappy, so tonight might be different. Pray for me oh world, pray for me!!!!