Everyday hero

I like any movie, but I love super-hero movies. Stories of people who can fly, stop time or throw balls of fire from their bare hands.

But when you think about it – all Christians should be super heroes. We have the power to read peoples thoughts, to see into the future, to heal the sick or even wake up the dead.

If you think about it…

And with the super hero task comes an arch enemy. And we have one.

And with great power comes a weakness. And we have that too. One that all heroes have. I’m not talking about kryptonite or lack of strength or that they can lock you into a prison made out of plastic because all you have power over is metal. It’s the weakness of mind. Doubting your purpose on this earth, doubting your abilities.

And I guess it’s hard to be a hero when you’re not wearing a tight fitting outfit with a big S on it.

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What to do on a Saturday OR How to make a Western blot

So now that I’m writing so much about my colleagues and the fascinating job I have as a researcher, you might wonder what it is that I actually do. The thing that occupies most of my time is doing Western blots. They should not be confused by Northern or Southern blots where you study RNA and DNA. Instead, Western blotting is a way to visualize proteins that you for example extract from cells.

Doing a Western blot (in one day)

1. Make gels. (these were prepared in advance and wrapped in wet tissue to keep them from drying)

2. Plan your Western. Sit down and really think about in what order you want to put your samples. Or how to put 16 samples in a gel that fits 14…

3. Load the samples into the gel. Easy enough…

4. Run the gel on 200V for: 10% gel=one hour, 12% gel=70min and 15% gel= one and a half hours. The proteins are negatively charged and should therefore travel towards the positive side.

5. Remove the gel from the case and put it next to a membrane so that the proteins will be transferred from the gel onto the membrane. The proteins are still negatively charged so be sure to put the membrane towards the positive side. Run on 40V for one and a half hours.

6. After the proteins have been successfully transferred you put the membrane in milk powder to prevent the antibodies you are using later on from binding to unspecific proteins. The red and blue stripes you see on the membrane is a ” ladder” which gives you the length of your proteins. (The milk powder we use is Valio’s, but if your group is low on cash I guess Pirkka does an equally good one.)

7. After blocking for one hour you can add the primary antibody. This will bind specifically to the protein of interest. You keep the membrane in TBS-T buffer together with the antibody for one hour on a “belly dancer”. The membrane is now put in those black boxes that you see on my lab table.

8. After washing 3*5min in TBS-T you add the secondary antibody. The primary one could have been produced by a rat, a mouse, a rabbit or even a goat, so depending on this you add anti-rabbit or anti-mouse or so on. One microliter from our stock is enough.

9. After this it’s very important that you thoroughly wash the membranes to get rid of any excess antibody, say 4*10min in TBS-T.

10. The secondary antibody was tagged with HRP so when adding  a developing substance the antibodies will light up and your proteins will be visible as fluorescent bands. Takes about 1-5min.

11. Then you take your membranes to the developing room, cover them with a regular Kodak film for 15 seconds up to 5 min, this have to be done in total darkness. Then you put the film in the developing machine and wait. Soon you’ll see the result as small black bands on the film – those are now your proteins.

12. Then you stare at, sorry- analyze your result and try to figure out what went wrong…

As you might have noticed, anyone could do a Western and sometimes I don’t think you’d need a six year long education to analyze the results either…

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