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Thank you for answering my enquiry. The conditions used for the Western Blotting in my lab are outlined below. The lysates are prepared in 4% SDS – so every single protein is extracted. As you can see from the attached .ppt file the control antibody worked fine – unfortunately the LSAMP antibody failed to produce the expected result. I am not going to spend more time on testing the antibody in question and want either to get a refund for the item or to get the rabbit polyclonal version of the antibody - ab95063.
P.S. On your website there is no image of the ab89719 antibody probed against an endogenously expressed LSAMP protein – usually this means that such tests either were not performed or proved to be unsuccessful.
The WB conditions:
The tissue lysate with confirmed LSAMP expression (15 μg of total protein per lane) was loaded onto a 4-12% Bis-Tris gel. Proteins were transferred from the gel onto Hybond-C nitrocellulose membranes. The membranes were blocked with 4% (v/v) nonfat dry milk for 1.5 hours prior to incubation with primary antibodies in TBS containing 0.08% Tween 20 (TBS-T) . The dilutions of primary antibodies were 1:500. Following the 4 hour incubation at RT, the membranes were washed with TBS-T. The primary antibodies were visualized with ECL peroxidase-conjugated secondary antibodies.
Asked on Nov 23 2012
Thank you for taking time to answer our questions and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
I apologize for the inconvenience and am pleased to offer you a free of charge replacement with ab95063 or a refund in compensation.
Thank you for your cooperation. I look forward to hearing from you with details of which resolution you would prefer.
Answered on Nov 23 2012