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Thank you for your reply.
I haven't try 2ug/ml of this ab.
The lot number is definitely GR65923-1. To prepare my samples, I diluted 2ul of mouse plasma in 60ul of 1X SDS loading buffer which contains freshly added beta-mercaptoethonal. I loaded 20ul of the diluted sample for each lane in 15% SDS-PAGE gel. Before loading, all the samples were boiled for 10min. I transferred the samples to a PVDF membrane from Millipore for 1h. Then blot it with 5% milk in TBST O/N at 4 degree on shaking. Next day, the membrane was incubated with 1ug/ml ab in 5% milk(you suggested using 1-2ug/ml in the data sheet) for 3h at RT on shaking. Washed with 1X TBST for 5mins X 3. Incubated with anti Rb -IgG HRP from Santa Cruz for 1h at RT on shaking. Washed as before.ECL.
We don't have much samples left and still need them to do other experiments. If you could agree with the replacement, I would appreciate it.
Asked on Feb 23 2012
Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab46819.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.
Answered on Feb 23 2012