Get Answer: Fafah Level Medical Question Guide

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Fafah, a level 400 Medical Laboratory Science student of Baldwin College performed immunohistochemical localization of oestrogen receptor on formalin fixed human breast tumours by an indirect method (using protocol A below) but the positive control failed to show. PROTOCOL A No Reagent Time/mins Xylene 5 Xylene 5 Absolute ethanol 5 Absolute ethanol 5 90 ethanol 3 80% ethanol 3 70% ethanol 3 Rinse in tap water 3 Heat sections in citrate buffer PH=6.0 using a pressure cooker 5 Rinse in tap water. 3 3 % hydrogen peroxide 5 Wash in PBS-2×. 5 Add 10% Bovine serum Albumin 20 Wash in PBS-2×. 5 Add primary antibody raised in mouse (1 in 1000 dilution) 60 Wash in PBS. 2×. 5 Add HRP-Labelled secondary antibody raised in rabbit (1 in 200 dilution) 30 Wash PBS.2×. 5 Add 3-amino-9-ethylcarbazole (AEC) 5 Rinse in tap water. 1 Counterstain with Mayer’s haematoxylin. 2 Rinse in tap water 5 70% ethanol 3 80% ethanol 3 90% ethanol 3 Absolute ethanol 5 Absolute ethanol 5 Xylene 3 Xylene 3 Mount with DPX STUDY THE PROTOCOL HE USED AND ANSWER THE FOLLOWING QUESTIONS 1. Give the specific names of the primary and secondary antibodies he used. 2. Draw and label a diagram to represent the immunohistochemical staining reaction. 3. What is the reason for step 9 in the staining protocol? 4. Explain the mechanism behind step 9 in the protocol. 5. Mention one (1) other method that ca

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