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Indirect ELISA test

ELISA test

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3 replies to this topic

#1 lilywong1991



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Posted 14 April 2015 - 08:35 PM

In the indirect ELISA test the enzyme-linked antibody will attach to ?


A)the patient antigen
B)the variable region of the patient antibody
C) the constant region of the patient antibody
D) the wall of the microtiter well
E) known antibody

Edited by lilywong1991, 14 April 2015 - 08:36 PM.

#2 mdfenko


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Posted 15 April 2015 - 05:17 AM

what do you think?


we'll tell you if you're correct.

Edited by mdfenko, 15 April 2015 - 05:20 AM.

talent does what it can
genius does what it must
i used to do what i got paid to do

#3 Zoe Winslet

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Posted 15 June 2016 - 06:55 PM

The indirect detection method adds a labeled secondary antibody for detection on the basis of direct ELISA and it is the most popular ELISA format. Antigen is passively attached to wells by incubation. After washing, antibodies specific for the antigen are incubated with the antigen. Wells are washed and all bound antibodies are detected by the addition of anti-species antibodies covalently linked to an enzyme. 

Be best yourself!

#4 Zagami Francesco

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Posted 08 August 2016 - 06:32 AM

Whole Toxoplasma Antibodies Research (WTAR)

Coated plate : reconstitute lyophilized Toxoplasma gondii antigen RH strain from peritoneal exudate from inoculated mice (Diagnostic Pasteur cod. 52721) with 2 ml of dH2O obtaining a concentration of 109 tachyzoites in 2.0 ml. Mix in order to obtain homogeneous suspension and aliquote in tubes with 0.4 ml of suspension (2.0x108 tachyzoites/0.4 ml) that is stored at -20 °C. Mix 0.4 ml of suspension with 0.6 ml of carbonate/bicarbonate buffer (35 mM sodium hydrogen carbonate and 15 mM sodium bicarbonate) at pH 9.6 obtained a tachyzoites concentration of 8x107 per ml. Dispense 100 µl of the final suspension in microplate well having antigen optimum concentration of 0.8x107  tachyzoites per well. Seal plate with adhesive plastic, and incubate overnight at 4 °C. After washing the wells with PBS containing 0.05% Twee-20 for 3 times, invert microplate and shake out the wash solution from the wells and dry. Use immediately or place the coated plate in plastic bag containing a drying agent (silica gel) and store at 4 °C resulting stable for 8 months.


Dispense in wells 100 µl of 1:100 diluted sera (1% BSA in washing solution) and incubate for 1 h at 37 °C. After another wash, dispense 100 µl of 1:1000 diluted peroxidase-conjugated rabbit or goat anti-human IgG and incubate for 1 h at 37 °C. Wash again and dispense 100 µl of TMB substrate.

This is the first immunoenzymatic assay realized by myself in 1993. Use this basis for elaborate a personal ELISA test.   

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