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Thread: Effect of EDTA on commercial antibodies

  1. #1 Effect of EDTA on commercial antibodies 
    Forum Sophomore M_Gabriela's Avatar
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    Hello!

    Im new in this forum. Im from a spanish speaking country (Argentina) so I apologize in advanced if I write something incorrectly.

    I would like to know if EDTA, being a chelating agent, inhibits the function of commercial antibodies (for example an antibody against a tag of a recombinant protein)

    Could EDTA function as a denaturing agent? What should I look for to understand the effects of this reagent on antibodies?

    Thanks in advanced

    Regards,

    Gabriela.


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  3. #2  
    Forum Masters Degree mat5592's Avatar
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    We use EDTA as an anticoagulant in the clinical lab all the time, including tests within the blood bank. Antigen typing uses commercial antibodies to determine a patient's rbc phenotype, so I would guess any inhibitory effect is negligible or we would be using one of the several other anticoagulant options for these tests.


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  4. #3  
    Forum Sophomore M_Gabriela's Avatar
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    Hi, thanks for answering!

    So, I will be honest with you. I, accidentally, dilute an alicuot of a very expensive antibody in EDTA 0.45 mM, instead of glycerol. I dont know how it happened, but it happened. I dont do this, actually this never happened to me before. I hope I wont get fired when they find out, hehe.
    So, as it is an antibody againt HA tag, and we purify recombinant protein with HA tag, I was going to do a western blot to check the function of the antibody.
    But for what Ive seen, EDTA is used 1-1.5 mg/mL as anticoagulant and the concentration in my tube is much more higher tan that.
    And also, I cant do the western without them watching so they are going to ask and I will have to tell them before I have the results...

    bye!!
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