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Serum 抗体 Purification Kit (Protein A) (ab109209)

Submit a review Q&A (4)References (1)

製品の概要

  • 製品名

    Serum Antibody Purification Kit (Protein A)
    Antibody Purification キット 製品一覧

  • 製品の概要

    Serum Antibody Purification Kit (Protein A) (ab109209) is prepared by coupling highly purified protein A to agarose beads and can therefore be used to purify IgG fractions from both serum and ascites fluid.


    The antibody is captured on the resin and unwanted substances are removed by a simple wash procedure. The purified product is then eluted and neutralized.


    The components of ab109209 are fully compatible with our Lightning-Link® Antibody Conjugation Kits and our Oligonucleotide Conjugation Kit. However they are not compatible with our nanoparticle Antibody conjugation kits (Gold, Latex, Europium and Magnetic). To purify antibodies for use with our nanoparticle conjugation kits, please use our Antibody Purification Kit - Nanoparticles (ab204909).


    ab109209 is not suitable for goat antibody purification. Please refer to the protocol booklet for more information about suitable and unsuitable species antibodies.

  • 特記事項

    This product is manufactured by Expedeon, an Abcam company, and was previously called AbSelect™ A Serum Antibody Purification System. 863-0500 is the same as the 3 tests size. 863-0030 is the same as the 1 test size.

    ab109209 contains 1-3 columns, and each can purify up to 20mg of antibody per run.

    The volume of sample required will depend on the host species (See Table below):

    Species Normal range IgG(mg/ml) Suitable Volumes for Product(ml)
    Rabbit 12 - 15 1.3 - 1.7
    Human 7 - 23 0.9 - 2.9
    Mouse 2 - 5 4 - 10
    Sheep/Goat 18 - 24 0.8 - 1.1
    Rat 5 - 7 2.9 - 4
    Ascites Fluid 0.5 - 5 4 - 40

     

     

製品の特性

参考文献 (1)

ab109209 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab109209 は 1 報の論文で使用されています。

  • Volgers C  et al. Budesonide, fluticasone propionate, and azithromycin do not modulate the membrane vesicle release by THP-1 macrophages and respiratory pathogens during macrophage infection. Inflammopharmacology 25:643-651 (2017). PubMed: 28528362

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1-4 of 4 Abreviews or Q&A

Question

I want to purify 500 ul of rabbit serum and do you think such a small amount would work ok with this kit as I was worried about diluting my antibody so much. Or could you recommend an alternative?

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Answer

It is true that in the protocol we recommend using 1.3-1.7ml of rabbit IgG.

0.5ml volume is therefore too low to use with this purification kit, so we would recommend diluting the antibody in a binding buffer to get a volume of 1.3ml and continuing with the protocol as normal . We do not see any reason why diluting the antibody further would affect their downstream process’ as there is an antibody concentration step at the end the protocol.

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Question

Dear Abcam team, I used your product (ab109209) for IgG purification from sheep serum and plasma, but I couldn't obtain the product enoughly. Actually I know that my serum and plasma include the specific antibodies against my specific organism in the high titers by IFAT. I tested the purified liquid for detection the antibodies by IFAT. However there was no reaction even in the low titers. I couldn't undertand why the purification was not successful. Would you kindly consider the problem? is the reason be able to related with Protein A? This work is very important for me because I will use it for immunoscreening of cDNA library. If you can comment the problem in a short time I would be happy. Thanks in advance. Ferda Sevinc, Professor, Turkey

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this purification kit.

I fully understand your concerns and it is regrettable the result have not been successful on this occasion. I am sorry to confirm that sheep antibodies do not have any affinity to protein A, so I would not expect this kit to work for sheep samples. In the previous email, advice was provided to confirm that the system for this kit is based on a Protein A purification and that the performance of the kit will therefore be both species and sub type dependent.

I would recommend a protein G kit would be more suitable for sheep antibodies, for example:

ab128751 Serum Antibody Purification Kit (Protein G)
https://www.abcam.com/Serum-Antibody-Purification-Kit-Protein-G-ab128751.html (or use the following: https://www.abcam.com/Serum-Antibody-Purification-Kit-Protein-G-ab128751.html).

If you would like to try this kit instead, I can offer you a 5% discount and free shipping for this kit. Please quote the followingcodes:

5% discount: #####
Free shipping: ####

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

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Question

I would like to use this Serum Purification kit on an antibody that is 60 uL of Ascites fluid. Should I dilute this antibody to a volume of 5mL? Will the purified antibody be at an acceptable concentration for use with the HRP conjugation kit or will I need to concentrate it further?

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Answer

Thank you for your phone calls.

I have heard back from the lab and they recommend using the purification kit ab102784 for small volumes of tissue culture supernatant or ascites fluid (100-500 uL). When using this kit, it will be fine to dilute your 60 uL of ascites fluid to 100 uL with PBS.

The purified antibodies can then be used with our EasyLink conjugation kits. For optimal results, the concentration of the antibody should be between 0.5-5 mg/mL. You may still be able to conjugate the antibody at lower concentrations, but it will likely not be as efficient and may result in unlabeled antibody present in the solution.

If you switch to the purification kits for smaller volumes, the concentration will likely not be a problem, but if you dilute your antibodies for use with the TCS or Ascites purification kits, you may need to concentrate it before use with the EasyLink kits. This can be done by using our concentration spin columns ab102776 or ab102778.

Since you have already purchased the TCS and Ascites purification kits, I will be happy to arrange an exchange if you would like to switch to the kits designed for smaller volumes. Let me know if you need any other information or assistance.

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Question

I bought your product (ab109209) for purifying sheep IgG, but I have not used it yet. The product can be used to purify IgG fractions from both serum and ascites fluid. I have a question about it. Can I use it for purifying IgG from sheep plasma? Please give me information in a shortest time. Greetings

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Answer

Thank you for your enquiry,

I can confirm that the testing oftheab109209 Antibody Serum Purification kit has been done on serum samples. Although there is no obvious reason why the kits should not work on plasma, I am sorry we presently have no data to confirm and guarantee this.

I would also advise that the system is based on a Protein A purification. The performance of the kit will therefore be both species and sub type dependent.

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

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