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For successful multiple labelling immunohistochemistry (labelling two or more antigens in the same cell) which of the following is true?Group of answer choicesthe secondary antibodies must be raised in the same species as the piece of tissue being studiedthe fluorescent labels must share the same excitation wavelength (ie: be activated by the same fluorescent light colour)the primary antibodies must be raised in the same species as the piece of tissue being studiedthe secondary antibodies must be raised in different speciesthe primary antibodies must be raised in different species

Question

For successful multiple labelling immunohistochemistry (labelling two or more antigens in the same cell) which of the following is true?Group of answer choicesthe secondary antibodies must be raised in the same species as the piece of tissue being studiedthe fluorescent labels must share the same excitation wavelength (ie: be activated by the same fluorescent light colour)the primary antibodies must be raised in the same species as the piece of tissue being studiedthe secondary antibodies must be raised in different speciesthe primary antibodies must be raised in different species

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Solution

The correct answer is "the secondary antibodies must be raised in different species".

Here's why:

In multiple labelling immunohistochemistry, you're trying to label two or more antigens (proteins) in the same cell. To do this, you use primary antibodies that bind to the specific antigens you're interested in. These primary antibodies are usually made in different species (like rabbit or mouse), so they have different "shapes" and can bind to different antigens.

Then, you use secondary antibodies that recognize and bind to these primary antibodies. The secondary antibodies are tagged with fluorescent labels, which allows you to see where the primary antibodies (and therefore the antigens) are located in the cell.

If the secondary antibodies were raised in the same species, they would all have the same "shape" and wouldn't be able to distinguish between the different primary antibodies. This would mean you couldn't

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