Overview

  • Product name
    Antibody Concentration Kit
  • Product overview

     


    Antibody Concentration Kit ab102778 allows for the quick and easy concentration of antibodies and proteins using a microcentrifuge-based method. The kit can also be used to reduce the concentration of many unwanted additives often found in antibody formulations such as azide, glycine or tris.


    Antibodies are sometimes only available at low concentrations and often contain low molecular weight substances that interfere in labeling reactions with enzymes, biotin, streptavidin and fluorophores. ab102778 can be used to avoid the impact of this.


    The Antibody Concentration Kit method utilizes a simple spin column to easily and quickly remove excess buffer from the antibody thereby providing a more concentrated antibody solution. The Antibody Concentration Kit also allows the experimenter to perform a simple buffer exchange to transfer the antibody.


    The components of Ab102778 are fully compatible with our Antibody conjugation kits however they are not compatible with our GOLD Antibody conjugation kits. To concentrate antibodies for use with our GOLD conjugation kits, please use our Gold antibody concentration kit (ab204911).


     

  • Notes

    Concentration of Antibody Solution

    1. Add antibody to spin cartridge.
    2. Spin for 1 to 3 minutes* in a microfuge at max speed of 15000g to reduce the buffer volume in the spin cartridge to between 50 and 100ul. It is advisable not to spin the antibody dry as reconstitution of the antibody will be difficult and significant antibody loss and degradation may occur.
    3. Repeat steps 1 to 2 as many times as is necessary to process the entire antibody to the desired concentration. It may be necessary to discard the excess buffer collected in the collection tube between spins.
    4. Recover the concentrated antibody from the spin cartridge. NB. It is advisable not spin the antibody dry as reconstitution of the antibody will be difficult and significant antibody loss and degradation may occur.

     Buffer Exchange Using Spin Column Assembly

    1. Add up to 0.5ml antibody to spin cartridge.
    2. Spin for 1 to 3 minutes* in a microfuge at maximum speed to reduce the buffer volume to 100ul.
    3. Discard the excess liquid in collection tube.
    4. Add 400ul conjugation buffer to the antibody in the spin cartridge.
    5. Spin for 1 to 3 minutes* in a microfuge at maximum speed to reduce buffer volume to 100ul.
    6. Discard the excess liquid in collection tube.
    7. Repeat steps 4 to 6 at least 5 times to exchange antibody buffer.
    8. Recover antibody from the spin cartridge.

     *Spin times will vary depending on buffer composition, starting volume and centrifuge speed.

    Note Each cycle leads to a reduction in the concentration of low molecular substances. By performing as many as 5 repeat steps the concentration of small molecules such as glycine and Tris will be reduced 2500 fold. However, the concentration of proteins such as BSA will be unchanged.

     To remove unwanted proteins see ab102784.

    Storage of Antibody

    Store at 4°C. Other storage conditions (e.g. frozen at -70°C may also be satisfactory). The sensitivity of any particular antibody to freeze thaw should be determined by experimentation on small aliquots.

    Test for Protein

    Wherever possible protein values should be determined using an absorbance at 280nm. For an IgG using a 1cm light path an OD280 of 1.0 is equal to an antibody concentration 0.714mg/ml.

    When using Bradford type reagents, it is important to use an IgG standard curve. The absorbance generated by this type of reagent is dependent on the protein used. For example using a BSA standard curve to determine the protein concentration of an IgG solution will result in a two fold under estimate of the IgG concentration.

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 3 units
    Conjugation Buffer 1 unit
    Spin Cartridge/ Collecting tube assembly 3 units
  • Research areas

Images

Protocols

References

This product has been referenced in:
  • Röper JC  et al. The major ß-catenin/E-cadherin junctional binding site is a primary molecular mechano-transductor of differentiation in vivo. Elife 7:N/A (2018). Read more (PubMed: 30024850) »
See 1 Publication for this product

Customer reviews and Q&As

21-26 of 26 Abreviews or Q&A

Answer

Thank you for contacting us. This kit (ab102778) is definitely meant to concentrate antibodies before labelling reactions. However, the process of labelling following the antibody concentration has only been done with our EasyLink conjugation kits, and we don’t have evidence of conjugation otherwise. https://www.abcam.com/index.html?pageconfig=resource&rid=13148&source=pagetrap Unconjugated fluorochromes can be removed by column chromatography or gel filtration. However, it is not always necessary to follow any method for this purpose since the fluorochromes may be removed during the washing steps as most of them (~99%) would be conjugated. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us. Unfortunately, we do not offer other concentrations of the antibody than the 0.25 mg/ml provided. However, we have antibody concentration kits available for the customer to concentrate ab24115. These are: ab102776: Click here (or use the following: https://www.abcam.com/index.html?datasheet=102776). ab102778: Click here (or use the following: https://www.abcam.com/index.html?datasheet=102778). To our knowledge, ab24115 has not been tested in EMSA yet. Therefore, I can offer a discount off a future purchase if you buy ab24115 now, test it in EMSA and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab24115 in EMSA. I will then send a discount code. This code must be issued before purchasing ab24115 so please wait for my reply before ordering. 2. Purchase ab24115 either by phone, fax, or online (www.abcam.com). 3. Test it in EMSA 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab24115 turns out to be unsuitable for EMSA, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.  

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Answer

Thank you for your interest in ab102778. Based on the description of your needs from this product, I would like to suggest an alternative product that would be more suitable. I would like to recommend the Antibody Tissue Culture Supernatant Purification kit. One column - ab109206 or 3 columns - ab109207. Each column can purify up to 50ml (5mg of antibody). I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Question
Answer

To sterilise the concentration tubes for kit 861-0010 we would recommend 70% ethanol, this will not harm the membrane. These concentrators are not suitable for autoclaving. I hope this is helpful. Please contact us again if you need any further information.

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Question
Answer

Thank you for contacting us with your question. My colleague who you previously spoke to is out of the office but I will be happy to help you. According to the production lab of ab102778, the concentration kit is suitable for removing glycerol from the antibody buffer. I hope this information is helpful, but please let me know if you have any further questions.

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Answer

Thank you for contacting us. For purifying from sera, we recommend ab102783 (1 purification) or ab102784 (3 purifications): Click here (or use the following: https://www.abcam.com/index.html?datasheet=102783). Click here (or use the following: https://www.abcam.com/index.html?datasheet=102784). The amount of antibody you recover can be checked by absorbance at A280 or with a protein assay such as BCA (bicinchoninic acid) or a Bradford assay. Please do not hesitate to contact us if you need any more advice or information.

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21-26 of 26 Abreviews or Q&A

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