Does anyone know if it's possible to detect immunoglobulins (preferably IgE) specific for Syphacia sp. (mouse pinworm), in serum?
To clarify, we wish to confirm the presence (or absence) of infection with Syphacia sp. from serum samples (from mice).
Preferably, we are interested in commercially available ELISA-kits.
Thanks in advance.
hi
If you not found commercially Elisa kit .
you can:
1- lyase syphacia sp by sonication in PBS 1X + Protease inhibitor cocktail (sigma)
2- measure Protein concentration by BCA method
3- set up ELISA test by
L1 = Coating = syphacia sp lysate ( such as 10 ug/ml ) ( 2 hour in 37c)
L2 = Blocking overnight with skim milk 3%
L3 = primary antibody = Titer mouse Serum from 1/100 to 1/32000 ( Normal mouse serum as negative control & Infected mouse as positive control )
L4 = secondary antibody = goat or rabbit anti mouse IgE - HRP conjugated
good luck
https://www.novusbio.com/products/ige-antibody_nb7533
Hello,
I think you may set up your ELISA platform as a specific ELISA kit may not be available for that.
In this case, the procedure described by @Reza Hadavi is . However here is a suggestion to increase the efficiency of the procedure (Step 3):
L2: Blocking overnight (at 4°C) with 5% BSA or skim milk.
L3: Use 1% BSA or Skim milk to dilute the primary antibody.
Thank you both, Reza and Kokuivi - I really appreciate that!
Would any of you be able to provide the adult worm, preferably sonicated? Or are you aware of anyone who works with this worm under controlled circumstances, that I could contact?
I would of course pay the shipment of this.
Best, Sebastian
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