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

11-20 of 26 Abreviews or Q&A

Answer

Thank you for your enquiry.

I have found 2 antibody concentration kits in our catalog, ab102776 (1 column)and ab102778 (3 columns). Could you confirm that one if these is the correct number?

For both these kits, I can confirmthey work using a size exclusion filter. The liquid that goes through the filter into the tube below will not contain the antibody; only molecules below a certain weight will go through the filter. The liquid that is left on top of the filter (in the spin cartridge)will contain the antibody. You can pipette out the liquid left in the spin cartridge, this containsthe antibody. there are no further steps required:

1. Add antibody to spin cartridge.
2. Spin for 1 to 3 minutes* in a microfuge at max speed of 15000 g to reduce the buffer volume in the spin cartridge to between 50 and 100 ul. It advisable not 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 advisable not spin the antibody dry as reconstitution of the antibody will be difficult and significant antibody loss and degradation may occur.


If you want to change the buffer the antibody is in, then I suggest to use the second protocol provided on the datasheet, for example if you want to put the antibody into some conjugation buffer ready for conjugation. Again, the antibody will remain in the liquid above the filter in the spin cartridge. You can spin through several lots of the newbuffer in order to wash away the original buffer.

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


I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for contacting us and your interest in our products. Kate us currently away from the office so I will try to help in your query.

Since your last enquiry, we have addedtwo protein G purification kits which you may want to consider:

Antibody purification kit (Protein G) https://www.abcam.com/Antibody-Purification-Kit-Protein-G-ab128747.html

and Serum antibody purification kit (Protein G) https://www.abcam.com/Serum-Antibody-Purification-Kit-Protein-G-ab128751.html

Both of these kits use protein G to purify antibodies and should be compatible with your sheep antibody. If you have any further questions regarding either of these products please do let me know and I will try to assist you further.

Read More

Answer

Thank you for contacting us. I have attempted to contact you by phone but thought I might communicate this information by email as well. I have reviewed of our catalog for mouse monoclonalAnti-Integrin beta 3 antibodies for you. I have also spoken with the lab regarding additives in the buffers of these products which might interfere with conjugation of HRP. After this I can say that AB110131 and AB7167 are the best options. However, each does need to be processed to remove stabilizers and preservatives which will interfere with HRP. At this time we do not offer any other alternatives which do not have these additives. To remove the preservatives, such as sodium azide, a buffer exchange/purification will need to be performed. OurAntibody Concentration Kit ab102778 (https://www.abcam.com/Antibody-Concentration-Kit-3-columns-ab102778.html)can remove these additives quickly and efficiently. The ‘clean’ antibody may then be conjugated using our EasyLink HPR conjugation kits (https://www.abcam.com/index.html?pageconfig=resource&rid=13148,https://www.abcam.com/EasyLink-HRP-Conjugation-Kit-3-x-10%C2%B5g-HRP-ab102892.html). We currently, until 31 May 2012, have a promotion whereas you can receive a free EasyLink conjugation kit with purchase of a primary antibody by using the code: These products are covered by our Abpromise guarantee. We provide scientific support, replacement or refund should this product not perform as indicated on the datasheet. More information on our Abpromise may be found at the following link: 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. Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

Read More

Answer

Thank you for your enquiry.

I can confirm that ab102778 Antibody Concentration Kit has a 10 kDa MW cut off filter, so it should besuitable to concentrate the Fab fragments inquestion.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for contacting us. We can sometimes arrange custom sizes and concentrations for bulk orders, and my colleagues at mailto:sales@abcam.com would be happy to put together a quote for you if you are looking to purchase 10 or more vials. For smaller quantities, you could use our concentration kits ab102778. These kits contain 10 kDa cutoff filters that are suitable for use with a protein as large as ab101823. I hope this helps, please let me know if you need any additional information or assistance.

Read More

Answer

Thank you for contacting us.

I'm afraid I can't disclose the composition of the conjugation buffer (because this is proprietary information) but I can tell you that is fully compatible with our conjugation kits.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

I'm glad to hear it. If you do have any further questions please do not hesitate to contact me again.

Until then, I wish you all the best with your experiments.

Read More

Question
Answer

I am sorry that the protocol details were not explained clearly enough. I will try to explain as best I can, if it does not make sense please let me know and I'll try to rephrase.

You would take your antibody (concentration of 0.36 mg/mL), you are hoping to achieve a concentration of at least 1 mg/mL to use it with theconjugation kit. In order to do this, you could concentrate all the antibody you will be provided with:

100 ug of antibody at 0.36 mg/mL, gives you 277.7 uL in total. This therefore needs to be concentrated by about a factor of 3 to get the desired concentration. You would therefore load the antibody into the spin cartridge. Spin (start with no more than 1 minute at no more than 15,000g), this will give you an idea of how quickly the solution will be removed. Continue to spin (checking after every minute or less or centrifugation) until you have removed ˜185 uL of solution. You should therefor be left with ˜92 uL of antibody at ˜ 1mg/mL.

If you would like to concentrate only a portion of the antibody, the same process can be followed but reducing the volume added and the volume discarded. Be very careful however to not let the antibody come to complete dryness (try not to reduce the antibody volume to below50-100 uL).

I hope this has helped to clarify the procedure. If you have any further questions, please do not hesitate to contact me again.

Read More

Answer

Thank you for contacting us yesterday.

Inow have some more input from the lab regarding the compatibility of the Anti-c Abl antibody [8E9](ab3227) with the R-Phycoerythrin Conjugation kit (ab102919). The lab have said that they recommend using antibody concentrations of 1-4 mg/mL, as this generally give the optimal conjugation results. With the antibody being at 0.2 mg/ml, if used with the R-Phycoerythrin Kit, we would expect to not obtain satisfactory results as the reactive chemicals would be too dilute.

We would therefore recommend that the antibody be concentrated prior to carrying out the conjugation procedure. This can be done using the antibody concentration kit, ab102778. It is worth having a look at the FAQs relating to the conjugation kits we offer which can be accessed from the link below:

https://www.abcam.com/index.html?pageconfig=resource&rid=13156

The antibody ab3227, once concentrated at least 5x would contain a higher concentration of BSA than ideal (concentration of BSA would be at least 1 %, recommended to be below 0.5%). It would therefore be advisable to also use the purification kit (http://releaseday.abcam.com/Antibody-Purification-Kit-Protein-A-ab102784.html) to remove the BSA.

I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

Read More

Answer

Thank you for your enquiry.

I can confirm that the concentration kits ab102778 and ab102783 are designed to concentrate antibodies and perform simple buffer exchanges. The kits are based on a10KDMCO filter. Therefore,theywill work perfectly well with sheep antibodies.The filters in the spin column are designed so that about 97% of the antibody should be recovered. The only difference is the size of the kit: ab102778 has 3 purification columns, ab102783 has one. I can recommend to review the online datasheets for further information

Another consideration is whether you want to conjugate the antibody. In this case, the antibody buffer should not contain BSA as this can inhibit conjugation. Because the molecular weight ofBSA is greater than10KD it will be concentrated alongside the antibody with this kit. If your antibody contains other proteins such as BSAthat you don't want, I can suggest to try a protein G purification for sheep antibodies. I am sorry we do not have any protein G purification kits in the catalog at the moment.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

11-20 of 26 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up