Paraoxonase (PON1) ELISA kit - (Jan/15/2014 )

Good day to all!

 

I am relatively new in the field of immunology as I have just transitioned into a new project from my previous one that is on qPCR.  I intend to investigate on the enzyme Paraoxonase in human plasma which is implicated in many diseases associated with the human liver.  I am looking for an ELISA kit that is specific to PON1 detection, preferably Sandwich ELISA due to experiment-related reasons.  Actually i was looking for an ELISA kit that is used to detect Paraoxonase 1 activity but that will be reserved for another day.  For now, it stays at enzyme detection only.

 

So I was just googling for a Sandwich ELISA kit to buy for my experiment.  I have to be clear on this so I would like to know, when it is shown as this:

Product Name    Paraoxonase-1 (PON1), ELISA Kit  Also Known As    Human Paraoxonase-1 (PON1) ELISA Kit  Product Gene Name    PON1 elisa kit

 

What does the "Human" part of Human PON1 ELISA kit refers to?  Is this kit used for detecting human PON1?  There are other PON1 ELISA kits with "Hamster", "Monkey", "Fish", "Rabbit", etc listed on the website.  So does this mean these ELISA kits are dedicated to detecting these animal-specific PON1?  As far as immunology is concerned, what I can gather (months after I have left school) is that if order to detect a protein, specifically, an enzyme, you need an antibody, in this case a primary PON1-antibody which is needed to bind to the antigen (PON1).  The primary antibody is then bound by secondary antibody that is then bound by the enzyme HRP that is conjugated with a substrate that is most probably a fluorescent probe and detection is thus made possible by analysing the fluorescence emitted using, say, a microplate reader.

 

In this case, I want to do a Sandwich ELISA which means I would need a capture antibody, primary antibody, secondary antibody, and a HRP-conjugated fluorescent probe.  The capture antibody, as the name suggests, is the ab used for capturing the enzyme and "lock" it in place for the primary ab to bind and subsequently the secondary ab and the HRP-conjugated probe yes?

 

Please do correct me if I have made any mistakes in my clarification.  Any additional info will be very much appreciated.

 

THANK YOU!

the species given in the kit name is the species from which the pon1, against which the primary (and capture) antibody are made, comes.

 

this wikipedia page can answer many of your questions: elisa

mdfenko on Wed Jan 15 13:47:21 2014 said:

the species given in the kit name is the species from which the pon1, against which the primary (and capture) antibody are made, comes.

 

this wikipedia page can answer many of your questions: elisa

Thank you for the prompt reply.  May I also clarify that in Sandwich ELISA, the reason why the clone numbers for the capture Ab (primary antibody) and the detecting Ab (secondary HRP-conjugated antibody) need to be different is to allow the detecting Ab to bind to a different site on the antigen before adding the fluorescent substrate for measuring the fluorescence.  If the clone numbers of both monoclonal antibodies are the same then the ELISA kit would not work yes?

yes, you require 2 clones which bind to different epitopes of the protein (unless the protein has multiples of the epitope).