Antibody Storage

 

Upon Receipt

• The prolonged activity of antibodies is best ensured by proper storage and handling practices. These can help to extend their activity by months. Please see below for general advice on antibody storage and handling. Please note: always consult the manufacturer’s data sheet for specific instructions.
• Upon receipt of the antibody, initiate the process by centrifuging it at 10,000 x g for 20 seconds to draw down the solution trapped in the vial threads. Subsequently, aliquot the antibody into low-protein-binding microcentrifuge tubes. This aliquotting procedure serves to mitigate damage caused by repeated freeze/thaw cycles, which can otherwise denature the antibody and lead to the formation of aggregates, diminishing its binding capacity. Additionally, aliquotting minimises the risk of contamination associated with multiple pipetting from a single vial.

Conditions

• It is best for the aliquots to only undergo freezing and thawing once, with the remaining quantity stored at 4°C. Generally antibodies are best stored at -20°C. Always verify the specific requirements for the antibody from the manufacturer's datasheet as the temperature may differ depending on the specific product.
• The size of aliquots should be determined based on typical experimental usage, with a minimum volume of 10 μL to mitigate the impact of evaporation and antibody adsorption onto the storage vial's surface.
• For the initial one to two weeks after receipt, storage at 4°C is generally acceptable, but adherence to datasheet recommendations is essential. To prevent freeze/thaw damage, avoid using a frost-free freezer and position antibody vials in an area with minimal temperature fluctuations.

Cryoprotectants

• It is common to use glycerol as a cryoprotectant at a concentration of 50%. This lowers the freezing point to below -20°C and thus helps to prevent damage from freeze/thaw. It is important to validate the antibodies’ stability when under this condition. Storing glycerol solutions at or below -80°C is not recommended as this is below the freezing point of glycerol. Please ensure that the glycerol used is sterile and free from bacteria.
• For conjugated antibodies, additional precautions are required due to their more complex nature. Conjugated antibodies should be stored in dark vials or otherwise protected from exposure to light which can impact their activity. Fluorescent conjugates are especially susceptible to photo-bleaching and thus should be protected from light sources throughout the experiment.
• Enzyme-conjugated antibodies should be handled with care to preserve their enzymatic activity and antibody binding capacity. It is advised not to freeze these antibodies; instead, they should be stored at 4°C unless a cryoprotectant is present, and the antibody's stability has been verified for long-term storage at -20°C.

Preventing Bacterial Contamination

• To safeguard against bacterial contamination, sodium azide may be introduced to the antibody solution at a concentration not exceeding 0.02%(w/v). The presence of sodium azide is indicated in the datasheet. However, caution should be exercised, most pertinently, when working with live cells or during in vivo experiments as sodium azide may be toxic to a number of organisms.
• Sodium azide should be absent when staining or treating live cells with antibody solutions and should be avoided entirely when dealing with HRP-conjugated antibodies. There exist a number of methods by which sodium azide may be removed; such as ultrafiltration and gel filtration.

Are you or your research group performing Parkinson’s-related research? Reach out to Biorbyt’s scientific support team here to schedule a consultation and see how Biorbyt reagents may support your team’s needs.