UA Biophysics:Protocols:Erythromycin

Erythromycin acts by inhibiting elongation at the transpeptidation step, specifically aminoacyl translocation from the A-site to P-site by binding to the 50s subunit of the bacterial 70s rRNA complex.

STOCK SOLUTION

Erythromycin is used in the biophysics lab at a 2 ug/ml. Stock solutions are at 1 mg/mL

PROTOCOL (STORAGE)

According to desired amount of antibiotics, calculate the amount in grams that has to be weighed.

For example, for 20mL of 25mg/mL of Erythromycin:

25mg/mL*20mL=500mg

1. Weigh the Erythromycin
2. Erythromycin has to be diluted in EtOH (molecular grade)
3. Divide in aliquots of 500μL
4. Store at -20 °C

Erythromycin is light sensitive. Keep tightly closed in light-resistant containers. This product is stable in solution at 37°C for 3 days. Stock solutions should be stored at 2-8°C.

USAGE

For culture plates, allow media to cool to 55 °C before adding antibiotic to a final concentration of 2 μg/ml.

For Example, for 300 mL of media at a final concentration of 30µg/mL:

C_i V_i = C_f V_f

34mg/mL* V_i = 0,03mg/mL*300mL

V_i = 9mg/(34mg/mL)

V_i = 0,264 mL

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