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小鼠抗体Purification试剂盒(ab128745)

Submit a review Q&A (6)References (2)

概述

  • 产品名称

    小鼠抗体Purification试剂盒
    参阅全部 Antibody Purification 试剂盒

  • 种属反应性

    与反应: Mouse

  • 产品概述

    Commercially available antibodies often contain substances (e.g. BSA, glycine, tris, azide) that interfere in labeling reactions with enzymes or fluorophores. ab128745 quickly removes these contaminants. It can also be used to purify antibodies from crude samples such as ascites fluid or immune serum. The mouse antibody to be purified or cleaned up ideally is in a volume of 100 µL to 0.5 mL. 20 µg to 150 µg of antibody can be purified in each run.


    The method involves capture of the antibody on a mouse resin and the removal of unwanted substances by a simple wash procedure, which is carried out in a standard microfuge. The purified product is then eluted and neutralized.


    Note: Binding strength of Bovine IgG to the Mouse resin is negligable.


    Antibodies purified using the Mouse Antibody Purification Kit 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. To purify antibodies for use with our nanoparticle conjugation kits (Gold, Latex, Europium and Magnetic), please use our Mouse Antibody Purification Kit - Nanoparticles ab204910.

  • 说明

    This product is manufactured by Expedeon, an Abcam company, and was previously called AbSelect™ Mouse Antibody Purification System. 830-0010 is the same as the 3 tests size. 830-0005 is the same as the 1 test size.

     

性能

  • 存放说明

    Store at +4°C. Please refer to protocols.
  • 组件 1 tests 3 tests
    ab273959 - Additional Collecting Tubes 4 vials 12 vials
    Binding Buffer (10X) 1 x 200µl 1 x 200µl
    Elution Buffer 1 x 1500µl 1 x 1500µl
    ab273960 - Mouse Resin 1 x 400µl 3 x 400µl
    Neutralizer Buffer 1 x 500µl 1 x 500µl
    ab273958 - Spin cartridge/ collecting tube assemblies 1 unit 3 units
    Wash Buffer 1 x 12ml 1 x 12ml

  • 研究领域

文献 (2)

发表研究结果有使用 ab128745？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab128745 被引用在 2 文献中.

  • Shaw HA  et al. Multicomponent Vaccines against Group A Streptococcus Can Effectively Target Broad Disease Presentations. Vaccines (Basel) 9:N/A (2021). PubMed: 34579262
  • Shu S  et al. An Immunosuppressive Effect of Melanoma-derived Exosomes on NY-ESO-1 Antigen-specific Human CD8+ T Cells is Dependent on IL-10 and Independent of BRAFV600E Mutation in Melanoma Cell Lines. Immunol Invest 49:744-757 (2020). PubMed: 32799717

客户评价及客户问答

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1-6 of 6 Abreviews or Q&A

Question

ab128745: binds which mouse isotype and subisotype?
does it bind IgA as well?

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Answer



The Mouse Antibody Purification Kit was developed to purify mouse IgGs and is suitable to purify any mouse IgG. This product was never tested on IgAs, although we do not think it will be as efficient as it is for IgG purification.

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Question

Customer saw peaks at 224 in all 4 elutions, but did not see peak at 280 nm. She followed the protocol using 150ug of antibody. She mentioned that adding the elution buffer formed a thicker, coagulated substance within the columns. Do you have any suggestions?

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Answer

Based on the description of the thicker, coagulated substance it sounds like the antibody precipitated in the low pH elution buffer. If this is the case then the precipitated protein will not go through the spin column, hence the absence of an absorbance at 280nm in the eluted fractions.
Would you mind sharing the information about the specific antibody that you were trying to purify? Is there any information on how it reacts to different pH levels? With additional information we may be able to come up with an alternative approach.

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Question

Do your antibody purification kits (for mouse-- protein G) from crude matrices (like sera) work for antibody isotypes, other than IgG (e.g. IgA, D, E, M)?

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Answer

The antibody purification kits that we carry, such as ab128745 and ab128747, will only recognize and purify IgG. For other classes such as IgA, IgD, and IgM, we have class-specific antibodies, for instance ab124716, which could be used to immunprecipitate those classes of immunoglobulin. Please let me know if you have any questions.

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Question

Customer is interested in ab41825, but wonders if we could label this one for him. Suggested ab102849 to do it himself to save money.
Will the 1% BSA interfer with the labeling process? Unfortunately, yes. ab128745 is necessary (or a pull down is perfomed within the lab).

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Answer

Vielen Dank für Ihren Anruf.

Leider sind 0,5% BSA das Maximum, dass das Labeling-Kit toleriert. Daher wäre entweder eine IP notwendig, mit der Sie den Antikörper von BSA aufreinigen notwendig, oder aber Sie benutzen ein weiteres Kit priori dem eigentlichen Labeling Kit.

Zusammenfassend lässt sich sagen dass Sie die folgenden Produkte von uns zur Konjugation erwerben können:

https://www.abcam.com/Cytokeratin-8--18--19-antibody-2A4-ab41825.html (or use the following: https://www.abcam.com/Cytokeratin-8--18--19-antibody-2A4-ab41825.html).

https://www.abcam.com/EasyLink-Alkaline-phosphatase-Conjugation-Kit-1-x-100g-AP-ab102849.html (or use the following: https://www.abcam.com/EasyLink-Alkaline-phosphatase-Conjugation-Kit-1-x-100g-AP-ab102849.html).

https://www.abcam.com/Mouse-Antibody-Purification-Kit-ab128745.html (or use the following: https://www.abcam.com/Mouse-Antibody-Purification-Kit-ab128745.html).





Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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Question

Customer kindly contacted us regarding the concentration of ab76424, lot GR-107769. Additionally, she is performing a HRP conjugation of the antibody and is running into issues with the 50% glycerol in attempting to remove sodium azide.

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Answer


As we discussed, our studies have found little effect on the conjugation chemistry with up to 0.05% BSA and up to 0.1% glycine in the antibody storage buffer. Thus, I think you do not have to worry about those components. However, as you know the 0.01% sodium azide will inhibit HRP activity, and as you mentioned the 50% glycerol is making it hard to remove the sodium azide.

Antibody purification kit if you are going to stay with ab76424:
Protein A for rabbit antibodies ab102784: https://www.abcam.com/Antibody-Purification-Kit-Protein-A-ab102784.html
Protein G for other species ab 128747: https://www.abcam.com/Antibody-Purification-Kit-Protein-G-ab128747.html
For mouse antibodies ab128745: https://www.abcam.com/Mouse-Antibody-Purification-Kit-ab128745.html

Previous customers have contacted us with issues purifying antibodies with high concentrations of glycerol, and we found the following solution proved successful:
" When using the antibody purification kit the presence of glycerol in the antibody buffer will make it more viscous. This means that it will require more G force to pass through the spin column. It is possible to buffer exchange the antibody it just means the customer will have to spin for much longer. Alternatively, a simpler solution would be to dilute the antibody. This would reduce the glycerol’s concentration, which in turn will make the buffer less viscous and easier to spin out. You can then use the same spin column to concentrate the antibody back to its original concentration and volume, after having performed the buffer exchange."

Additionally, we have our EasyLink conjugation kits, including HRP conjugation, listed through the link below. Our EasyLink conjugation kits are capable of fully successful conjugation with glycerol concentrations of up to 50%. We are currently running a promotion for an extra 20% off our EasyLink kits:
EasyLink Kits: https://www.abcam.com/index.html?pageconfig=resource&rid=13148#enzymatic
Easylink Promotion:https://www.abcam.com/index.html?pageconfig=resource&rid=15389&source=pagetrap&viapagetrap=easylink123
Below is a list of compatible and incompatible buffers with our EasyLink conjugation reactions:
https://www.abcam.com/index.html?pageconfig=resource&rid=13156#11

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Question

Thanks for the information, so for example for ab16838 which comes from tissue culture supernatant and has 1%BSA and is only 500 ul, would you recommend the Mouse Antibody purification kit, https://www.abcam.com/Mouse-Antibody-Purification-Kit-ab128745.html, rather than the Mouse TCS purification kit, https://www.abcam.com/Mouse-TCS-Purification-System-ab128749.html, since the volume is so small? I was wondering if the ab128745 kit could be used on TCS?

Thanks,

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Answer

Thank you for your reply.

You are spot on about using the Mouse antibody purification kit, ab128745, to remove the BSA, sodium azide, glycine, tris, and other molecules that may be present in a volume of 100-500ul. The kit is compatible with ab128745 and other TCS antibodies. ab128749 is listed as "TCS" in its title for purifying antibody from large, non-purified, TCS volumes.

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

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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