Anti-Glutathione antibody [D8] (ab19534)

"It is technically challenging and we do not recommend it for quantitative analysis.

The best method developed so far is that of Sun and Kalloniatis, J Comp Neurol 494:686 -703, 2006.

We advise to consider this when doing cryosections immunocytochemistry for small molecules like glutathione:

1. All our small molecule antibodies are designed for post embedding immunocytochemistry using plastic resin. Cryo imaging is not a technology we recommend, but we understand some users wish to use it anyway. It is possible, but the user must appreciate that this turns a quantitative method into a qualitative one.

2. Small molecule detection requires some glutaraldehyde fixation. We recommend 0.05% for about 60 minutes followed by a buffer wash and conventional cryoimmunocytochemistry with standard detergent penetration. The choice of visualization is up to the user, recognizing that glutaraldehyde increases background fluorescence, mostly in the the green (Alexa 488) range. This is less of a problem for Cy3, Cy3.5 and other very red dyes. It is not a problem for HRP/DAB. Typically the 1:100 primary dilution we recommend may be a little strong for cryo and lower levels of 1:200-1:500 may be necessary, depending on how much glutathione there is in the tissue (e.g. kidney and liver have much higher levels than skeletal muscle, heart or nervous system).

3. All cells have some glutathione ranging from 100 uM to 10 mM depending on types. This means that in a superposition image, there will be a very high glutathione "background." Thus we recommend confocal imaging to minimize this superposition effect."

I hope thisadditional information is helpful and wish godd luck with your research.

I have been informed by the lab that the conjugation of KLH to glutathione was through the thiol group.

As requested I issued a credit note. The credit note ID xxxxxx.

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion.
I appreciate the time your customer has spent on these experiments and would be pleased to arrange a replacement with a different lot number or a credit note in compensation.

I look forward to hearing from you with details of how you and your customer would like to proceed.

We have 2 antibodies that react with conjugated glutathione and 2 that are designed to react with free glutathione. I have looked at these products and it doesn't appear any of them have been specifically tested for reactivity to glutathione as a thioether. So they may or may not react with glutathione as a thioether.