Ampicillin Stock Solution

Ampicillin Stock Solution: A Comprehensive Guide

Ampicillin, a broad-spectrum beta-lactam antibiotic, is frequently used in research and clinical settings to inhibit bacterial growth. Rather than preparing a fresh solution each time it's needed, creating a concentrated ampicillin stock solution offers significant advantages in terms of time efficiency, accuracy, and minimizing waste. This article provides a detailed guide to understanding, preparing, and using ampicillin stock solutions.

1. Understanding Ampicillin and its Solubility

Ampicillin is a water-soluble antibiotic, meaning it dissolves readily in water. However, the concentration at which it dissolves completely depends on factors like temperature and pH. The generally accepted solubility of ampicillin in water at room temperature is approximately 100 mg/mL. This means you can dissolve a maximum of 100 milligrams of ampicillin powder in 1 milliliter of water. Exceeding this limit will result in undissolved ampicillin powder remaining in the solution, leading to inaccurate concentration and potentially ineffective treatment. Choosing the right solvent (water, in this case) and ensuring thorough mixing are crucial steps in preparing a stable and accurate stock solution. A higher concentration stock solution necessitates the use of a higher volume of solvent.

2. Calculating Ampicillin Stock Solution Concentration

Determining the required concentration of your ampicillin stock solution is the first crucial step. This depends entirely on the final concentration needed in your experiment or application. For example, if you require a final concentration of 100 µg/mL ampicillin in your cell culture and you need 10 mL of this solution, you need to calculate the amount of ampicillin needed from your stock. Let's assume you are making a 1000x stock solution (i.e., 100 mg/mL). To obtain 10 mL of 100 µg/mL solution, you would use the following formula:

C1V1 = C2V2

Where:

C1 = Concentration of the stock solution (100 mg/mL or 100,000 µg/mL)
V1 = Volume of the stock solution needed (unknown)
C2 = Desired final concentration (100 µg/mL)
V2 = Desired final volume (10 mL)

Solving for V1: V1 = (C2V2)/C1 = (100 µg/mL 10 mL) / 100,000 µg/mL = 0.01 mL

Therefore, you would add 0.01 mL of your 100 mg/mL stock solution to 9.99 mL of the appropriate solvent (e.g., cell culture media) to achieve your desired final concentration. This illustrates the practicality of using a concentrated stock solution – you are only adding a small volume to achieve your final concentration.

3. Preparing the Ampicillin Stock Solution: A Step-by-Step Guide

The process of preparing an ampicillin stock solution is relatively straightforward but requires meticulous attention to detail. Here’s a step-by-step guide:

1. Weighing the Ampicillin: Accurately weigh the required amount of ampicillin powder using an analytical balance. Remember to account for the desired concentration and the final volume.

2. Dissolving the Ampicillin: Add the weighed ampicillin powder to a suitable sterile volumetric flask containing a portion of sterile water. Gently swirl the flask to dissolve the powder completely. Avoid vigorous shaking to prevent foaming. Add more sterile water to bring the volume up to the calibration mark.

3. Filter Sterilization (Recommended): Filter sterilize the solution using a 0.22 µm filter to remove any bacterial contaminants. This is especially crucial for applications involving cell cultures or in vivo studies.

4. Storage: Store the ampicillin stock solution at -20°C in appropriately labeled aliquots to prevent repeated freeze-thaw cycles. Ampicillin solutions are generally stable for several months under these conditions.

4. Factors Affecting Stability and Shelf Life

Several factors can influence the stability and shelf life of an ampicillin stock solution. These include:

Temperature: Higher temperatures accelerate degradation. Freezing at -20°C is crucial.

pH: Ampicillin is more stable at a neutral or slightly acidic pH.

Light Exposure: Ampicillin is susceptible to degradation by light. Store the solution in amber or opaque containers.

Repeated Freeze-Thaw Cycles: Avoid repeated freeze-thaw cycles as they reduce the potency of the antibiotic.

5. Applications of Ampicillin Stock Solutions

Ampicillin stock solutions find widespread applications in various fields:

Microbiology: Used in bacterial growth inhibition assays, antibiotic susceptibility testing, and preparation of selective media.

Cell Culture: Frequently used to select for antibiotic-resistant cell lines or eliminate bacterial contamination in cell cultures.

Pharmaceutical Research: Used in preclinical studies to evaluate the efficacy and safety of new drugs.

Clinical Settings: Prepared as a concentrated stock for intravenous or intramuscular administration (though typically prepared by pharmacists in clinical settings, not researchers).

Summary

Preparing an ampicillin stock solution is a vital technique in many research and clinical settings. Accurate calculation, proper weighing, careful dissolving, and appropriate storage are key to maintaining the solution's integrity and potency. Remembering to always prioritize sterility, especially when preparing solutions for sensitive applications like cell culture, is absolutely critical. This detailed guide provides a comprehensive understanding of the process, allowing researchers and students to confidently prepare and utilize ampicillin stock solutions effectively and efficiently.

FAQs

1. How long is an ampicillin stock solution stable? A properly prepared and stored ampicillin stock solution at -20°C can remain stable for several months, but it's always advisable to check for any signs of degradation (e.g., cloudiness, precipitation) before use.

2. Can I prepare a larger volume of ampicillin stock solution? Yes, you can prepare larger volumes, but ensure you have the appropriate equipment (e.g., larger volumetric flasks) and maintain sterile conditions throughout the process.

3. What should I do if my ampicillin stock solution appears cloudy? A cloudy solution indicates potential degradation or contamination. Discard it and prepare a fresh solution.

4. Can I freeze-thaw my ampicillin stock solution multiple times? No, repeated freeze-thaw cycles degrade ampicillin, reducing its efficacy. Aliquot your stock solution into smaller volumes for single use.

5. What are the safety precautions I need to take when handling ampicillin? Always wear appropriate personal protective equipment (PPE), such as gloves and eye protection, when handling ampicillin powder or solutions. Avoid direct contact with skin and eyes. Dispose of the solution according to your institution's guidelines.

Search Results:

Ampicillin (Amp) Stock Preparation - Benchling This protocol describes the preparation of an Ampicillin stock solution. The concentration of this stock solution is based on the weight of anhydrous Ampicillin. Note: Typical stock solutions for this antibiotic in selectable media are made at 50mg ml-1 and are used at a final concentration of 20-40 μg ml-1 for stringent plasmids and 100 μg ...

bio.display » Basic Protocols - Tyrell Ampicillin stock solution. Add 5ml 96% ethanol to 500mg of amicillin in a sterile vial and stir. keep in deep freezer, -20°C. After doing so, disperse into 1ml or 0.5ml aliquots for storage. This will result in an ampicillin stock of 100mg/ml. The final concentrations we’re aiming for is 100µg/ml.

Ampicillin - OpenWetWare 9 Feb 2008 · Stock Solution. Typical concentrations of ampicillin are 50 μg/mL for low copy plasmids and 100 μg/mL for high copy plasmids. Stock solutions are typically at 100 mg/ml, so that 1 ml of antibiotic can be added to 1 liter of broth or agar. Stock solutions made in 50% alcohol remain liquid at -20°C and are easy to pipet.

Preparation of ampicillin stock solution in water - Laboratory Notes A 50 – 100 mg/ml stock solution of ampicillin is generally prepared in water. Depending on the need, the stock solution is diluted to the final working concentration. Commercially, ampicillin is available in three forms – Ampicillin anhydrous, Ampicillin trihydrate and Ampicillin sodium salt.

Ampicillin stock solution (100 mg/mL) - CSH Protocols Dissolve 1 g of sodium ampicillin in sufficient H 2 O to make a final volume of 10 mL. If sterilization is required, prewash a 0.45- or 0.22-μm sterile filter by drawing through 50-100 mL of H 2 O.

Ampicillin Stock Solution - goldbio.com Sterilize Ampicillin Stock through the prepared 0.22 μm syringe filter. Stock may be kept at -80°C for 1 year. Note: Ampicillin solutions degrade 13% after one week at -20°C.

Antibiotics Stock Solutions | Nicolas Kylilis Working Solution: Typical concentrations of ampicillin are 50 μg/mL for low copy plasmids and 100 μg/mL for high copy plasmids. Stock solutions are typically at 100 mg/ml, so that 1 ml of antibiotic can be added to 1 liter of broth or agar.

Ampicillin, 100 mg/mL, Ready Made Solution, A5354, Sigma-Aldrich Order Ampicillin solution, a semisynthetic derivative of penicillin, from Sigma-Aldrich. Ideal for Cell Biology and Biochemical research.

Ampicillin Preparation and Selection Guide - abo Stock solution: An ampicillin stock solution can be prepared at a concentration of 100 mg/mL and should be stored at -20⁰C. 1. Add 1 g (1000 mg) of ampicillin to 10 mL of dH 2 O 2. Sterilize the solution using a 0.22 μm filter 3. Store solution in different aliquots at -20⁰C LB-ampicillin agar preparation: 1. Dissolve the following in 500 ...

Preparing Antibiotics Stock Solution and Ampicillin Agar Plates Add Ampicillin-Na or Kanamycin stock solution to an end concentration of 100 µg/ml Stir or shake accurately to get a homogenous antibiotics distribution! Pour about 25-30 ml in a 10 cm dish

Antibiotic stock solutions The stock solutions is filtered through a 0.2 m filter, aliquoted and stored at -20oC until use. The final working concentration of all antibiotics is 1:1000 of the stock solution. Pouring agar Plates 1. Wipe down the bench to with 70% EtOH or bleach. 2. Remove foil cover from LB agar flask but leave sponge. 3.

Ampicillin Stock Solution (1000×) - CSH Protocols Ampicillin Stock Solution (1000×) Prepare a 100 mg/mL ampicillin sodium salt (Sigma-Aldrich A9518) solution in H 2 O. Filter-sterilize and store for up to 3 mo at 4°C. © 2023 Cold Spring Harbor Laboratory Press

Ampicillin Stock Solution Protocol 100 mg/ml - GoldBio Sterilize Ampicillin Stock through the prepared 0.22 μm syringe filter. Stock may be kept at -80°C for 1 year. Note: Ampicillin solutions degrade 13% after one week at -20°C. GoldBio's …

Ampicillin - 10 mL (1,000 X Stock) - Cytographica 1) Dissolve 1 g of ampicillin into 10 ml of ddH 2 O. 2) Filter through a 0.22 µm filter to sterilize. 3) Aliquot and store at -20°C. 4) Use at 1:1000 dilution in LB or LB-Agar.

Ampicillin - PombEvolution Ampicillin is stored as a 1000X stock in freezer 7. Use in LB medium at a final concentration 100mg/L. Ampicillin is not stable at high temperatures, so don't forget to cool down your medium to <60°C before adding the ampicillin. Ampicillin stock solution (100 mg/mL) Dissolve 0.5 g of sodium ampicillin (in freezer) in 4 ml MilliQ water

Ampicillin Anhydrous Stock Solution - GoldBio "Stability of frozen stock solutions of beta-lactam antibiotics, cephalosporins, tetracyclines and quinolones used in antibiotic residue screening and antibiotic susceptibility testing."

Ampicillin Stock Solution Preparation and Recipe - novoprolabs.com Store the ampicillin in aliquots at -20°C for 1 yr (or at 4°C for 3 mo). In general, use carbenicillin for ampicillin selection, because it is more chemically stable. Working Conc. = 50-100 µg/mL.

Ampicillin Stock Solution - GoldBio Sterilize Ampicillin Stock through the prepared 0.22 μm syringe filter. Stock may be kept at -80°C for 1 year. Note: Ampicillin solutions degrade 13% after one week at -20°C.

ProtocolsAntibioticStockSolutions - Barrick Lab Ampicillin degrades quickly in both plates and stock solutions. Culture plates with Amp can be stored at 4°C for about 2 weeks. Stock solutions can be stored at 4°C for 2 weeks but can last as long as 4-6 months when stored at -20°C.

Protocol: Ampicillin (Amp) Stock (100 mg/mL) Recipe 13 Sep 2019 · Protocol: Ampicillin (Amp) Stock (100 mg/mL) Recipe Application: Making 100 mg/mL Amp stock solutions for use in Luria-Bertani (LB) liquid media or LB agar plates. Procedure: 1. Mix 2 g Amp powder into 20 mL of MilliQ H 2O. 2. Vortex until powder is thoroughly dissolved. 3. Sterilize using 50 mL syringe with syringe filter into 50 mL conical ...

Ampicillin Stock Solution