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Location: Home > Information Center > FAQs > Antibody preservation FAQs

Antibody preservation FAQs

Date: 2016-03-31 Author: Leading Biology Click: 1761

Antibody preservation FAQs


The activities and using effects of antibodies depend on preservation. Most antibodies activities can last for months, even years with properly storage. Please always store the antibodies correctly according to the recommendations on the datasheet.


1. Centrifuge at 12000 rpm for 1-5 minutes before open the tube cover for aliquotting and storage (If the antibody is less than 50 ul, please extend the centrifuge time to 5 minutes to ensure that all antibodies are dropped down) after receipt.


2. For most antibodies, freeze at -20°C or -80°C in small aliquots is the optimal storage condition.


For most antibodies, it’s sufficient to store at -20°C. There is no appreciable advantage to storing at -80°C.


Aliquotting minimizes damage due to repeated freezing/thawing, as well as contamination introduced by pipetting from a single vial for many times.


It is better to use up a aliquot in one experiment. Aliquots should be no smaller than 10 ul. The smaller the aliquot, the more the stock concentration is affected by evaporation and adsorption of the antibody onto the surface of the storage vial.


Aliquots should be frozen and thawed once, with any remainder kept at 4°C.


The antibody working fluid should be used up the day after you prepared. Avoid keeping the solution at 4°C for more than one day.


Never store the antibodies in the automatic defrost refrigerator. Try to keep the antibodies in the middle of the refrigerator rather than keep near the door.


3. In most cases, store at 4°C upon receipt of the antibody is acceptable for 1-2 weeks. If the antibodies is used in 1-2 weeks, it’s better to store at 4°C to minimize damage due to repeated freezing/thawing. Recommend to store the antibodies at -20°C or -80°C for a long time. It is important to follow the recommendations on the datasheet.


However, ascites fluid may contain proteases, and should be frozen as soon as possible after receipt. The antibodies would degrade if the ascites store at 4°C for a long time.


4. Transportation conditions for most antibodies: Recommend to transport at 4°C because general transport process would take 1-2 weeks. The main aim to transport at 4°C is to avoid the damage to the antibodies activity by repeated freezing/thawing (We will receive frozen antibodies if transport with dry ice. Thawing is needed to aliquot, which adds another round of freezing/thawing). Therefore, avoid transport with dry ice.


Transportation conditions for special antibodies:

Enzyme-conjugated antibodies: Enzyme-conjugated antibodies should be kept at 4°C. Avoid freezing because it will decrease the activity of enzymes, even inactivation .


Conjugated antibodies: All the conjugated antibodies should be stored in dark vials or wrapped with foil to shielded from light. Fluorescent antibodies in particular are sensitive to light and should be protected from light during all phases of an experiment.


IgG3 isotype: IgG3 isotype should be kept at 4°C. Do not freeze because this kind of antibodies tend to form aggregates during freezing/thawing (No matter storage or transportation), which will reduce the antibodies binding capacity.


Related information about antibodies storage


Add glycerol:

Some people will add 50% glycerol into antibodies to avoid repeated freezing/thawing because glycerol could lower the freezing point at -20°C. It’s useful for most antibodies. Solutions added with glycerol are not recommended to keep at -80°C because it has exceeded the freezing point of glycerol. Pay attention to aseptic operation and avoid contamination if you want to store the antibodies with glycerol.


Protein protectant:

High concentration proteins (1 mg/ml or higher) are not easy to degrade. That is the reason why proteins such as BSA are added into antibodies as stabilizers. On the other hand, the protein minimize the loss of antibodies absorbing to the wall. Don’t add stabilizers if you want to mark with antibodies because they will compete with antibodies to conjugate markers.


Use sodium azide:

Sodium azide is always added into antibodies to avoid microbial contamination (Final concentration is 0.02% (w/v)).


Avoid using sodium azide in the following conditions:

When antibodies is used to stain or treat live cells and in vivo research;


Sodium azide is toxic to most organic compounds while resisting microbes because it will cut off mitochondria cytochrome electron transportsystem.


Conjugate with antibodies that involve amine groups:

Sodium azide will interfere with any conjugation that involves an amine group. Thus, it is necessary to remove sodium azide before conjugation. Conjugated antibodies can be stored only in 0.01% sodium azide. Thimerosal (merthiolate) could be a substitute for sodium azide to protect antibodies which need to be conjugated.


Methods to remove sodium azide

Sodium azide can be removed by gel electrophoresis or filtration. IgG molecular weight is 150 kDa (IgM is 600 kDa), but the sodium azide molecular weight is only 65 Da. A filter film with a cut off at 14 kDa can remove sodium azide from the antibodies.


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