You have added IgG-sensitized red cells to the negative indirect antiglobulin test result of the antibody screen procedure. You
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As mentioned above, phosphoric acid has 3 pKa values, and after 3 ionization it gives 3 types of ions at different pKa values:
H₃PO₄(aq) + H₂O(l) ⇌ H₃O⁺(aq) + H₂PO₄⁻ (aq) pKₐ₁
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</span>H₂PO₄⁻(aq) + H₂O(l) ⇌ H₃O⁺(aq) + HPO₄²⁻ (aq) pKₐ₂
HPO₄²⁻(aq) + H₂O(l) ⇌ H₃O⁺(aq) + PO₄³⁻ (aq) pKₐ₃
At the highest pKa value (12.4) of phosphoric acid, the last OH group will lose its hydrogen. On the picture I attached, it is shown required protonated form of phosphoric acid before reaction whose pKa value is 12.4.
H₃PO₄(aq) + H₂O(l) ⇌ H₃O⁺(aq) + H₂PO₄⁻ (aq) pKₐ₁
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</span>H₂PO₄⁻(aq) + H₂O(l) ⇌ H₃O⁺(aq) + HPO₄²⁻ (aq) pKₐ₂
HPO₄²⁻(aq) + H₂O(l) ⇌ H₃O⁺(aq) + PO₄³⁻ (aq) pKₐ₃
At the highest pKa value (12.4) of phosphoric acid, the last OH group will lose its hydrogen. On the picture I attached, it is shown required protonated form of phosphoric acid before reaction whose pKa value is 12.4.