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Thanks for the response. I didn't see that before, but it was actually a typo on my part. What I should have written is that the 1:100 dilution of the 0.82 mg/mL stock gave a 820ug/dL solution (alternatively, an 8.2ug/mL solution as you indicated). Approximately 161.5uL of the 8.2ug/mL solution was then diluted to a total of 500uL (just a bit more than a 3-fold dilution), to give a final concentration of about 2.65ug/mL (or 265ug/dL), which was treated as the most concentrated sample for the standard curve. The remaining points were created by 1:2 serial dilutions of that sample. One note though... for the ˜3-fold dilution, we have been using the standard diluent supplied with the ALerCHEK ApoB ELISA kit. I could re-try the protein using a different diluent like PBS or would you have a suggestion for a better diluent? Thanks! On Thu, Jan 12, 2012 at 1:30 PM email@example.com wrote: We have an answer to your inquiry: Thank you for sending the kit catalogue number. Something I overlooked in your description of the problem was your lab's preparation of the ApoB. It is supplied at a concentration of 0.82 mg/ml, = 820 ug/ml. Your lab apparently diluted this 100-fold, according to your e-mail from October 21: Supplied at 0.82mg/mL in 200mM Tris, 10mM SDS, pH 8.25. Diluted 1:100 to 820ug/mL with 1x PBS, pH 7.4.
Asked on Jan 12 2012
Thank you for the clarification. I had a feeling it would not be that simple! I expect that the buffer supplied in the kit for diluting experimental samples is suitable for standards too. You might ask the kit supplier if their standard is equivalent to ab77859, though they may be reluctant to approve another company's material as a subsitute for their own standard. Our product is more specifically referred to as ApoB-100, as it is gel-purified to remove smaller ApoB lipoproteins, but I do not think this would make ab77859 incompatible with this kit.
Answered on Jan 12 2012