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    products/conjugation-kits/antibody-concentration-and-clean-up-kit-ab102778.pdf

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Antibody Concentration And Clean-Up Kit (ab102778)

  • Datasheet
  • Protocol Booklet
Submit a review Q&A (26)References (5)

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Antibody Concentration And Clean-Up Kit (3 columns) (ab102778)

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    Overview

    • Product name

      Antibody Concentration And Clean-Up Kit
    • Product overview

      Antibody Concentration And Clean-Up 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 And Clean-Up 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 And Clean-Up Kit also allows the experimenter to perform a simple buffer exchange to transfer the antibody into a more favorable buffer for conjugation.


      Antibodies concentrated using the Antibody Concentration And Clean-up Kit are fully compatible with our Lightning-Link® Antibody Conjugation Kits and our Oligonucleotide Conjugation Kit. However they are not compatible with our nanoparticle conjugation kits (Gold, Latex, Europium and Magnetic).


      To concentrate antibodies for use with our Gold and Magnetic Conjugation Kits, please use Antibody Concentration And Clean-Up Kit – Gold And Magnetic Nanoparticles (ab204911). To concentrate antibodies for use with our Latex and Europium conjugation kits, please use Antibody Concentration and Clean Up Kit for Latex & Europium (ab269965).


       

    • Notes

      This product is manufactured by Expedeon, an Abcam company, and was previously called AbSelect™ Antibody Concentration and Clean-Up Kit. 861-0010 is the same as the 3 units size.

       

    Properties

    • Storage instructions

      Store at +4°C. Please refer to protocols.
    • Components 3 tests
      ab275797 - Conjugation Buffer 1 x 15ml
      ab273914 - Spin Cartridge / Collecting Tube Assemblies 3 units
    • Research areas

      • Kits/ Lysates/ Other
      • Kits
      • Concentration Kit

    Images

    • Antibody Concentration And Clean-Up Kit (3 columns) (ab102778)
      Antibody Concentration And Clean-Up Kit (3 columns) (ab102778)

      Representative Antibody Concentration and Clean-Up Kit (3 columns) (ab102778).

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (5)

    Publishing research using ab102778? Please let us know so that we can cite the reference in this datasheet.

    ab102778 has been referenced in 5 publications.

    • Ochocka N  et al. Specialized functions and sexual dimorphism explain the functional diversity of the myeloid populations during glioma progression. Cell Rep 42:111971 (2023). PubMed: 36640350
    • Boggs AE  et al. Optimization, validation and initial clinical implications of a Luminex-based immunoassay for the quantification of Fragile X Protein from dried blood spots. Sci Rep 12:5617 (2022). PubMed: 35379866
    • Saitoh Y  et al. Improvement of hepatocyte engraftment by co-transplantation with pancreatic islets in hepatocyte transplantation. J Tissue Eng Regen Med 15:361-374 (2021). PubMed: 33484496
    • Calvani M  et al. Preliminary Study on ß3-Adrenoreceptor as Predictor Marker of Relapse in Ewing Sarcoma Patients. Biomedicines 8:N/A (2020). PubMed: 33066095
    • 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). PubMed: 30024850

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 26 Abreviews or Q&A

    Question

    I purchased your product above (concentration kit) to concentrate purified antibody. I would like to enquire if the columns can be re-used or are meant to be used once and discarded?

    Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Jul 10 2015

    Answer

    Thank you for your enquiry.
    The kit can be re-used up to 6 times. However, it can only be re-used with the same antibody. You could put wash buffer on its own through the column in between washes to clean it, but large particles will never get completely removed from the column so we would not recommend re-using the kit with other antibodies or proteins due to cross-contamination issues.
    I hope this information will be helpful. If you have any further questions, please do not hesitate to contact me.

    Read More

    Sam Washer

    Abcam Scientific Support

    Answered on Jul 10 2015

    Question

    Customer wants to purify mouse monoclonal IgG that is in PBS and azide. Which kit to use?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 04 2015

    Answer

    Thank you for your call and your patience while I communicated with my colleague in the lab.

    In response to your inquiry, it turns out that both options (ab102784 Antibody Purification Kit (Protein A) or ab102778 Antibody Concentration Kit ) will work for your antibody purification purposes. However it turns out that the simplest kits would be the Concentration kit.

    The spin columns provided in this kit allow a simple buffer exchange by ‘spinning away’ the existing buffer, and allowing this to be replaced by a more compatible buffer. As spinning the antibody dry is not best practice, as this can impact the antibody, the protocol states that this allows the azide to be reduced, as there will be a small amount of azide left in the little bit of buffer that remains. However if this process is repeated, each time, the amount of azide remaining will reduce more and more, therefore after enough spins, the azide concentration will effectively become zero. On the whole, this azide should be reduced to a suitable level almost after 1 buffer exchange, however we would recommend this is repeated further if the label of choice is HRP, as this has shown to be more sensitive. I hope you find this information helpful.

    Please let me know if I can offer any more help or technical advice. Good luck with your research.

    Read More

    Abcam Scientific Support

    Answered on Mar 06 2015

    Question

    I plan to concentrate 30-40kDa Fab solution with Antibody Concentration Kit (ab102778). May I ask the mechanism of this Kit? Will the small size molecules be passed through the filter during centrifugation? Will FITC (400˜Da) be passed through?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 25 2014

    Answer


    The Antibody Concentration Kit method utilises a simple spin column to easily and quickly remove excess buffer from the antibody thereby providing a more concentrated antibody solution. Only molecules smaller than 10 kDa will be pass through the filter. Therefore, FITC alone will pass through the filter, unless it is conjugated to a molecule larger than 10 kDa, such as the antibody that you are concentrating.

    Read More

    Guillaume Boucher

    Abcam Scientific Support

    Answered on Sep 25 2014

    Question

    I will try to conjugate the anti-4 Hydroxynonenal antibody (ab48506) to FITC by using EasyLink FITC Conjugation Kit, according to your suggestion. I read the antibody concentration kit datasheet and I was wondering if better instruction can be provided about the Buffer Exchange protocol. For instance, is there a spin speed that is ideal (an optimum spin speed) and how many minutes are required to reduce the buffer volume from 500 to 100 microliters?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 21 2013

    Answer

    Most benchtop microcentrifuges have a maximum spin of 14,000 - 15,000 RPM, and a maximum RCF of 18,000 - 21,000 x g. Anywhere in that range will be fine. You might want to test spinning just buffer without antibody, to see how long it takes to reduce 500 ul (or less) to 100 ul, but an antibody solution will likely take longer, since the antibody blocks the filter. You can always check every 30 seconds to see how much the volume has been reduced, taking notes. After the first spin, the subsequent spins should take the same amount of time, assuming you add conjugation buffer after each spin (for example, PBS) to bring the volume up to the original volume of the Tris-buffered antibody solution.
    The hydroxynonenal antibody ab48506 will be 50 ug in 500ul, a concentration of 0.1 mg/ml. You will want to concentrate the antibody with the same spin column by not adding more buffer after the last buffer exchange spin. Ideally, you want a concentration of at least 1.0 mg/ml for efficient conjugation.

    Read More

    Tom Ruyle

    Abcam Scientific Support

    Answered on Jun 21 2013

    Question

    I have a 3ml antibody sample to concentrate with ab102778. I would like to know what is the maximum volume of antibody I can add to the spin cartridge. Can I use the one spin cartridge multiple times to concentrate the same antibody sample?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 18 2013

    Answer

    The maximum volume of antibody that you can add to the spin cartridge is 0.5 mL. However, you are can use the spin cartridge multiple times to concentrate the 3 mL antibody sample.

    Read More

    Abcam Scientific Support

    Answered on Feb 18 2013

    Question

    Customer contacted us regarding the use of the antibody concentration kit (ab102778) to concentrate 1ml of a 30kDa Fab fragment solution into 50-100uL. Both antibody concentration and buffer exchange procedures are not yielding increased concentration - measure by nanodrop at OD280 before and after concentration protocols are performed.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 17 2013

    Answer


    The laboratory responded that the filter in the antibody concentration kit ab102778 should work fine with a 30KD fragment. It is possible that the filter could be damaged. It would help if you could measure the OD of the flow through. If the flow through has the same OD then there is a problem with the filter. If not, then there may be an issue with the OD measurements, perhaps a blanking issue or issue with the reader itself.

    Please let me know the results of the OD readings of the flow through. I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on Jan 17 2013

    Question

    I am searching for ab15272 without sodium azide.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 30 2012

    Answer

    Thank you for contacting us. We do not currently sell ab15272 without sodium azide, but we may be able to custom formulate the antibody for bulk orders of more than 10 vials. For enquiries about a custom formulation, please contact mailto:sales@abcam.com. For smaller quantities, you may be able to perform your own buffer exchange to significantly reduce the amount of sodium azide by using our concentration kits: ab102776 and ab102778. I hope this helps, please let me know if you need any additional information or assistance.

    Read More

    Abcam Scientific Support

    Answered on Oct 30 2012

    Question

    MWe rencently check your company has anti-human TNFRII clone MR2-1 which contain 0.02% azide.

    My question is can you make a azide free of anti-human TNFRII (MR2-1) for us?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 22 2012

    Answer

    Thank you for contacting us. We may be able to custom make this antibody in an azide-free form for bulk orders of 10 or more vials. If you are interested in a custom bulk purchase, please contact my colleagues at mailto:sales@abcam.com and they can help put together a quote and estimated availability time for you.

    Alternatively, for smaller purchases you could use one of our concentration and clean up kits (ab102778) to perform a buffer exchange and greatly reduce the concentration of azide in the antibody. I hope this helps, please let me know if you need any additional information or assistance.

    Read More

    Abcam Scientific Support

    Answered on Oct 22 2012

    Question

    Customer kindly contacted us as they had excessive antibody loss during buffer exchange and labeling. To help with their troubleshooting of this, they where hoping that we could explain the use of PEG and sucrose as buffer for this AB as compared with other stabilizers such as BSA. Is this product sticky so that during buffer exchange it may aggregate?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 12 2012

    Answer

    Thank you for contacting us regarding ab58555.

    I have contacted the lab on your behalf, the antibody would be fine as well in a different buffer containing another stabilizing agent e.g. BSA. However this product is provided in PEG and sucrose as was found the antibody was suffering less during lyophilization.

    The antibody itself is not stickier than other antibodies. It seems possible that the buffer with the PEG and sucrose is responsible of the low yield that you are getting after the conjugation. The buffer exchange method may need to be adjusted for improved performance.

    We would recommend use of one of our concentration kits for performing buffer exchange with this product. Our antibody concentration and clean up kits, Ab102776 and ab102778utilizes a simple spin column to easily and quickly buffers. These should work in any buffer containing PEG and are covered by our Abpromise guarantee. Which guarantees scientific support, replacement or refund should any Abcam product not perform as indicated on the datasheet. More information about Ab102776, ab102778 and our Abpromise may be found at the link below.



    Ab102776:https://www.abcam.com/Antibody-Concentration-Kit-1-column-ab102776.html

    Ab102778:https://www.abcam.com/Antibody-Concentration-Kit-3-columns-ab102778.html

    https://www.abcam.com/index.html?pageconfig=abpromise

    I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

    Read More

    Abcam Scientific Support

    Answered on Oct 12 2012

    Question

    We are interested in conjugating our LIF antibody (see attached datasheet) with your easylink HRP kit (ab102891). I was wondering what kits we would need to order to successfully achieve conjugation. I wasn't sure if we needed to worry about purifying this particular antibody since it is already affinity purified and is in PBS/gelatin/azide. Also, I noticed your HRP kits recommend .5mg/mL - 5mg/mL for opitmal results but this antibody is only .2mg/mL. Would using one of the antibody concentration kits (ab102778) work to remove the small amount of azide and gelatin and also bring the concentration into optimal ranges for conjugation with the 3x100ug HRP kit? If you could let me know what materials we would need to achieve HRP conjugation with this particular antibody I would greatly appreciate it. Thanks so much!

    Read More

    Abcam community

    Verified customer

    Asked on Jul 13 2012

    Answer

    Thank you for contacting Abcam regarding our EasyLink conjugation kits.


    I have reviewed the product specs of the antibody you are using and the buffer formulation is compatible with the HRP kit ab102891. As you mentioned, the only modification I would recommend is to further concentrate the antibody for optimal results. I would recommend ab102778 for this purpose, as you have already suggested. You should not require any additional reagents to successfully complete the conjugation reaction.


    I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

    Read More

    Abcam Scientific Support

    Answered on Jul 13 2012

    1-10 of 26 Abreviews or Q&A

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