Overview

  • Product name
    Anti-Carboxymethyl Lysine antibody [CMS-10]
    See all Carboxymethyl Lysine primary antibodies
  • Description
    Mouse monoclonal [CMS-10] to Carboxymethyl Lysine
  • Host species
    Mouse
  • Tested applications
    Suitable for: ELISA, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    N epsilon (Carboxymethyl) Lysine (CML) conjugated to KLH.

Properties

Applications

Our Abpromise guarantee covers the use of ab30922 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA Use a concentration of 0.1 - 0.5 µg/ml.
IHC-Fr Use a concentration of 2 µg/ml.
IHC-P Use at an assay dependent concentration.

Target

  • Relevance
    N epsilon carboxymethyl lysine (CML or Carboxymethyl Lysine) is formed by the non enzymatic Schiff base reaction of glucose with proteins, followed by an Amadori rearrangement and oxidation that leaves only a carboxymethyl group attached to the lysine. The levels of CML adducts accumulate over time and have been used as an indicator of both serum glucose levels and oxidative protein damage. Elevated serum CML modified proteins have been associated with diabetes and may contribute to diabetic retinopathy, nephropathy and angiopathy.
  • Alternative names
    • CML antibody
    • N Epsilon (Carboxymethyl) Lysine antibody

References

This product has been referenced in:
  • Portal-Núñez S  et al. Adverse Effects of Diabetes Mellitus on the Skeleton of Aging Mice. J Gerontol A Biol Sci Med Sci 71:290-9 (2016). Read more (PubMed: 26386012) »
  • de la Maza MP  et al. Weight increase is associated with skeletal muscle immunostaining for advanced glycation end products, receptor for advanced glycation end products, and oxidation injury. Rejuvenation Res 11:1041-8 (2008). IHC-P . Read more (PubMed: 19086911) »
See all 2 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for providing those extra details. There are a few points which would warrant optimisation in order to obtain better results withthis antibody:


1. Storage


From the datasheet we recommend that this antibody be stored at -20°C on delivery. It may be thathaving stored it at +4°C may have reduced its activity.


2. Substrate coating


It may be that optimising the coating conditions would improve the signal observed. I would suggest trying incubation for2 hours at room temperature. I would also consider trying to use a carbonate buffer for coating, a recipe and description of use can be found here:


https://www.abcam.com/ps/pdf/protocols/Direct%20ELISA%20protocol.pdf


3. Blocking


Unless already doing so I would suggest blocking for2 hours at room temperature.


4. Incubation of primary antibody


Incubation at 37oC is not suitable for many antibodies. In the optimisation stages I would suggest using 2 hours at room temperature, or 4oC overnight can often produce stronger signals. If these conditions result in an observed standard curve, incubation at 37oC could then be attempted.


5. Secondary antibody


I could not find the exact concentration or recommended dilution of the secondary antibody being used however this can be crucial in your experiment. We would typically recommend a dilution in the region of 1/4000-1/10,000. This may yield more specific results.


I understand that you have been trying to get this ELISA to work effectively for some time and the effort you have gone to. I am therefore willing to offer a replacement with the antibody you want (ab27684). If you would still like to proceed with this please do let me know and I will have it arranged. In order to do this I will send the replacement directly to you, could you therefore provide your shipping and billing address as well as your institutions VAT number.


Many thanks for your assistance in this matter.

Read More

Answer

Thank you for contacting us.

We normally use heat mediated antigen retrieval in IHC-P. The extra information when we have, is always written as application note under the application. I have checked the data currently we have for this antibody and unfortunately we haven't tested this antibody in IHC-P in our own lab. The IHC-P application is added because of an Abreview which is also published online. I also have attached a publication which mentions successful use of this ab in IHC-P. In both case the heat mediated antigen retrieval method has been avoided.

I presume the heat might have damaging effect on the structure CML thereby giving false results. I therefore can suggest using enzymatic retrieval method or following the protocol as mentioned in the attached publication.

If you are interested I can provide you abreview discount code for using this antibody in IHC-P. Using the discount code you can avail a free antibody in return of an Abreview. Please click on the link for more details;

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

Let me know if you are interested in my offer.

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Answer

Thank you for contacting us and relaying the problems this customer has been having with ab30922.I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would appreciate it if you can confirm the following details:

1. how long after purchase was the antibody used and found to give low signals?

2. how was the substrate captured to the plate? Under what temperature, time?

3. how was the primary antibody incubated (temperature, time? with agitation?)

4. what wash steps were employed?

5. was there any sample concentration related increase in signal (ie increase with increasing concentration)? Could you please share the data obtained? This would greatly improve my understanding of the problem encountered.

6. which secondary antibody was used with the experiment (manufacturer and catalogue number would be most helpful).

I appreciate that alot of time has been spent in the lab trying to achieve good results with this antibody but by confirming these details I may be able to further conclude where the problem may be stemming from.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement (with an alternative antibody if requested) orcredit note.

I hope this information is helpful, and I thank you for your cooperation.

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Question
Answer

Thank you for contacting us.

Unfortunately we have no data about the cross-reaction of this antibody with Fructolysine.

Known cross-reactivities have been shown for Glyoxal and Glycolaldehyde. The antibody is preabsorbed for CEL and shows little binding to 3DG-modified protein.

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

I'm sorry I was not clearer in my explanation. Both ab30917 and ab30922 should be suitable for you experiment. The reason why we sometimes have what seems to be duplicates within our catalogue is that in order to make sure we have a continuous supply of an antibody which we outsouce, we also supply a back up, in case one of the suppliers is unable to continue providing us with the antibody. So if we are unable to continue selling ab30917 we would be able to provide ab30922 for our customers and vice versa. I hope this explanation helps. I wish you well in your experiments. If you have any further questions please do not hesitate to ask.

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Answer

Thank you for contacting us. The two antibodies (ab30917 and ab30922) are both mouse monoclonal antibodies directed against N Epsilon (Carboxymethyl) Lysine (CML). They have both been tested with ELISA, IHP (frozen sections and paraffin). They are of the same concentration (0.250 mg/ml) and have been purified in the same way (Protein G purified). The only difference is that the clones are different (ab30917 clone number CMS-10 and ab30922 and NF-1G). This means the antibodies come from different lines of hybridoma. This should have no effect on the performance on the antibody relative to the other one. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (intervertebral disc)
Specification
intervertebral disc
Fixative
Formaldehyde
Antigen retrieval step
Enzymatic
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10 %

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Verified customer

Submitted Jan 03 2007

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