Calcium gluconate is widely used in medical settings to treat conditions caused by low calcium levels, such as hypocalcemia, which may result from calcium deficiency, kidney failure, or thyroid surgery. It is administered intravenously to quickly address severe calcium deficiencies, which can lead to muscle spasms, cardiac issues, or tetany. In emergency situations, it is also used as an antidote for magnesium sulfate overdose and to counteract the effects of hyperkalemia by stabilizing the heart's electrical activity. Additionally, it is applied topically to treat burns caused by hydrofluoric acid, as it helps neutralize the acid and reduce tissue damage.

In veterinary medicine, calcium gluconate is used to treat hypocalcemia in pets such as dogs and cats, administered intravenously or orally under veterinary guidance.

In the food industry, it serves as a calcium supplement to fortify products like calcium-enriched milk or health supplements and improve nutritional value. Its use should be under medical or professional supervision to prevent side effects such as injection site irritation or hypercalcemia.