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Thank you for your helpful advice. I used Ponceau S on my most
recent blot and was able to visualize transfer of both my MW marker
and proteins in my cell lysate. It's certainly possible, however,
I could be getting over-transfer to some extent. I recently contacted
the lab that wrote the review for ab65166 on your website, and they
said that they performed a 16 h transfer at 20 V. With the new lysate,
my hope is to try both your one hour transfer conditions and the 16 h
transfer conditions and see if I see better transfer with either.
I'd also be open to trying higher MeOH and lower SDS content;
although, the aforementioned group apparently used a basic transfer
buffer with 20% MeOH and no SDS.
I'm not currently blotting for a loading control, but that is
a good suggestion. The best evidence I have for the success of my
WB protocol is that I could visualize Invitrogen's Precision
Plus MW marker after incubation with my secondary antibody.
In terms of lysing my own sample, I used the following reference to
prepare my lysis buffer (http://www.springerprotocols.com/Full/doi/
I certainly am open to try other lysis buffers but felt it might
be best to use a formulation that had been used successfully with
these channels before.
I very much appreciate the offer to send me another vial of ab30151.
From some initial reading, it looks like Nav1.6 expression increases
during maturation, so it's perhaps best to stick with the lysate
I'm already using. The order number for the original order is
I can't thank you all enough for your help so far. Your team
has been incredibly responsive, and I very much appreciate your
Asked on May 16 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 with the order number xxx. The estimated delivery date is xxx as the item is currently backordered. I hope this is not causing a big problem. I apologize.
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 May 16 2012