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Thank you for your reply.
This customer has tried some modifications and sent back the results.
Attached please find the data (4 pages).
He has tried to:
- Lyse sample with RIPA buffer.
- Heat sample 95C for 10 mins. (previously he heated the samples in 70℃ for 10min is due to the protocol of preparing sample from the Invitrogen kit; http://tools.invitrogen.com/content/sfs/manuals/nupage_minigel_qrc.pdf)
- Block with 5% BSA in TBST
- 1Ab dilution(1:1000) with 5% BSA, 4C O/N
- 2Ab dilution(1:5000) with 5% BSA in TBST at RT for 1Ab
It is mentioned in a study (attached) that LonP1 are expressed in human blood, and he therefore would like see the clear signal like the reference showed. Unfortunately, multiple bands were still detected.
Would you please help him check the data?
Thank you very much.
Asked on Feb 20 2012
Thank you for confirming the details.
The image you have kindlyattached is the same images you have previously sent.Is there no improvements at all. Could you share any new image you have?
Have you used lysates other than primaryfibroblasts? Are thesefibroblasts mouse or human?
Looking forward to hearing from you soon.
Answered on Feb 20 2012