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Author Topic: C-fos antibody Inactivated  (Read 3081 times)

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Offline pandanac

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C-fos antibody Inactivated
« on: December 29, 2011, 03:02:44 PM »
Hi All:

I have a big problem in my work.

1.I use IHC for staining C-fos protein in frozen brain sections.

2. Three weeks back, our refrigerator was turned off by a worker  who happened to be working in our lab for some construction  :( .
Before this fiasco took place... i was getting very good results...and fantastic staining... however..after this mess up I did  a test to see if the antibodies are working well..I did see some staining however it was not according to the exposure I had planned (Control, tactile and Visual exposure of rats...three stay in dark.. the control and tactile is killed in dark [objects are in the cage for the tactile exposure]...the other rat is exposed to light to stimulate the cfos.. as the rat would be seeing several objects[outside] in a controlled environment)
3. I saw the staining in the tactile to be more than the visual..which is quite contrary to the basic result i got..
4. The refrigerator is a 4 degree refrigerator and the antibody was a SC one.. and they had specified to store the Ab at 4 degree Centigrade.
5. I even had the Vecta Stain kit and the DAB kit....
6. The refrigerator was off for 14-15 hours..

My questions:

1. Is the antibody..inactive?(the results are as told above with a test)

2. Is the kits working properly?

3. Do the Ab work well even after such accidents?

4. The brain sections...have been in the freezer from Sept mid week... is it too long for them not to respond to a antibody..

5. If the Ab is totally inactive .... how is it working in tactile and very little in visually stimualted rats.. (I have used a section in the -8.00 Bregma.. which has a major part of visual area...)

Please tell me as soon as possible.... I am going crazy here.. and my entire 1 year work will come to a halt.. if this dosent work...

Please Help.!!!!

Thanks...!!!!


C-fos antibody Inactivated
« on: December 29, 2011, 03:02:44 PM »