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The order confirmation number is 1031347.
1) How much protein was loaded per lane of the gel?
I didn't measure protein concentration. Cells were cultured in a 6-well plate, and were confluent when harvested. Each sample was collected by adding 200ul 1xSDS sample buffer to one well of the 6-well plate. 20ul of the indicated sample (1/10 of the lysate) was loaded per lane of the gel.
2) What protease inhibitors were used, and was this a standard reduced and denatured Western blot?
I used mammalian protease inhibitor cocktail. Yes it was a reduced and denatured western blot.
3) What was used to block the membrane (i.e. milk or BSA)?
For this experiment I used 5% BSA.
4) What dilution of antibody was used?
Besides, I'd like to know more about the testing discount program. If the SCAP antibody (ab125186) works in human samples I would certainly test it on mouse or rat samples.
Asked on May 04 2012
Thank you for your reply and for the additional information.
I am sending a vial of ab86415 free of charge on the order ***. This antibody is in stock and should arrive on Monday. Please keep me updated about how this antibody works in your Western blots.
Since Ab125186 has not been tested in mouse or rat samples, I can offer a discount off a future purchase if you buy the antibodynow, test it in one of these speciesand submit feedback to us in the form of an Abreview. The discount would be worth1 free primary antibody.
If you are interested in this offer, please follow these steps:
1. Reply to this e-mail to let me know that you would like to proceed and test ab125186 in mouse or rat. I will then send a discount code.
2. Purchase ab125186 either by phone, fax, or online (www.abcam.com).
3. Test it in mouse or rat.
4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.
5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab125186 turns out to be unsuitable for mouse or rat, you will still receive the discount on your next purchase after your Abreview has been submitted.
Please let me know if you have any questions about this offer and I would be happy to help you further.
The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.
Answered on May 04 2012