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I have a plan to purchase ELISA kit (ab125299). I have two questions about ELISA kit.
Could you please let me know what kind of amine coated plate do you recommend (including company, number,,,) ?
When I measure absorbance, which OD do I need to use for measuring HIF-1a and PDK1 ? I will use FLUO star Omega (BMG LABTECH) instead of Odyssey.
Thank you for your time.
Asked on Apr 11 2012
1. The amine coated plates that we use are from BD Biosciences
BD PureCoat™ Microplates
96-well, amine 5
cat # 354717
Alternativley, you may be interested in the following product, which contains reagents (except antibody) required for ICE: https://www.abcam.com/ICE-In-Cell-ELISA-Support-Pack-ab111542.html. The Em/Ex for the IR secondary dyes are as follows: IRDye 800 (for PDK1, mouse) 778/794; IRDye 700 (for HIF1a, rabbit) 680/694.
2. Regarding the FLUO star Omega (BMG LABTECH): we have no experience with this reader and it will be up to the user to determine if in fact it is compatible with the IRDyes (based on their Em/Ex spectra above). From what I can see on the BMG LABTECH website, compatibility will be dependent on the users particular model (BMG lists Spectral Range as 240 - 740 nm or 240 - 900 nm).
Alternatively the user could try substituting their own dye-conjugated secondary antibodies that they are already certain work with their instrument. The PDK1 mAb is from mouse; HIF1 from rabbit.
I hope this is helpful. Please contact me again if you have any further questions.
Answered on Apr 11 2012