Goat Anti-Mouse IgG H&L (Cy5 ®) preadsorbed (ab6563)

Thank you for contacting us.

This antibody detects all mouse IgG, so it should work with both your IgG1 and IgG2 antibodies.

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

Thank you for your inquiry.

I used to perform many ELISPOTs by hand as you with plastic plates. The capture antibody was coated on the plates. The PVDF membrane bottom well plates areused when using a reader to analyze the results. The membranes are pushed through onto sticky tape and read in a reader with special software to identify spots.

I am not aware of a technique where the cells can be identified and sorted according to the production amount of protein X. I am assuming that protein X is secreted and therefore should be captured in the vicinity of the producing cell, but any slight movement of the plate will swirl the cells and make a identification impossible.

ab6563 is in general suitable as secondary to a mouse IgG1. But is is anti heavy and light chain. So it will pick up the light chain too. I suggest to choose an antibody specific to the heavy chain. I can recommend ab136127:

https://www.abcam.com/index.html?datasheet=136127 (or use the following: https://www.abcam.com/index.html?datasheet=136127).

This antibody is not specifically cross absorbed against IgE, but against Mouse IgM, IgG2a, IgG2b, IgG3 and IgA, pooled Human sera and purified Human paraproteins. ab136127 is tested in ELISA and therefore should also work in a ELISPOT setting as secondary.

I hope this information is helpful. Please do not hesitate to contact me again with any further questions in this matter.

Sorry for the delay in getting back to you.

I can now confirm that the product has been cross adsorbed against the species listed as showing minimal cross reactivity. This includesBovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rat, Rabbit or Sheep Serum Proteins.

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

Thank you for contacting us and sorry for the delay in getting back to you in regards to your query.

I have looked into the antibody you are interested in, the Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Cy5 ®), pre-adsorbed (ab6563), and can share some more detail about its purification.This productis prepared by initially using immunoaffinity chromatography using Mouse IgG coupled to agarose beads. This is then followed by solid phase adsorptions to remove any unwanted reactivates to serum of other species and extensive dialysis.

Cross reactivity of this antibody has been assessed and noreactionhas beenobservedwith Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Rat, Rabbitor Sheep Serum Proteins.

I am checking further with the source of thisantibody to ascertain if the antibody has been pre-adsorbed with all of these species of serum and will get back to you as soon as I have this information.

I am sorry for the delay and any inconvenience this causes you.

Thank you for contacting Abcam.

All of the information regarding purification of the antibodies in our catalogue is available on our website. If we do not specify that Protein A/G was used in the purification process then they were not used to purify the antibody.

I have looked through the list you sent and for some there is no purification notes, as is the case for ab3354 as it is supplied as an Ascites fluid but for ab3356, it does say that Protein G was used for purification purposes.

Please let me know if there is anything else I can help you with.

Thank you for your reply.

The information for how each antibody is purified is available on our website. Just go to the webpage for each antibody and the information is there. I have attached a document that shows you where to find that information.

If you have anymore questions after looking at the webpages, please let me know.

Thank you for contacting Abcam.

The antibody ab6563 was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities and extensive dialysis.

Please let me know if there is anything else I can help you with.

Thank you very much for your interest in our antibodies. The difference between these two secondary antibodies it that ab6563 is pre-adsorbed against 10 species (chicken, cow, goat, guinea pig, hamster, horse, human, rabbit, rat, sheep), while ab97037 is pre-adsorbed against only 8 species (chicken, cow, goat, horse, human, rabbit, rat, sheep). It will depend on your lysate, which one would be better suited. If you are using a human cell line for example ab6563 would be better suited. Although we have not tested these antibodies, we believe they would work and could be used in Western blot, provided you have the appropriate equipment for the detection of Cy dyes. Please do not hesitate to contact us or the manufacturer of your detection system for more information. Given this, I can offer a discount off a future purchase if you buy ab6563 or ab97037 now, test it in Western blot and submit feedback to us. It doesn’t matter whether the feedback is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free secondary antibody. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab6563 or ab97037 in Western blot. I will then send a discount code. This code must be issued before purchasing ab so please wait for my reply before ordering. 2. Purchase ab6563 or ab97037 either by phone, fax, or online (www.abcam.com). 3. Test it in Western blot. 4. Let us know the results, positive or negative, preferably via an email to technical@abcam.com (images are great if you have them!) similar to our Abreview system (which can be found here: https://www.abcam.com/abreviews.) 5. After the feedback is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any secondary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody turns out to be unsuitable for Western blot, you will still receive the discount on your next purchase after your feedback has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. Otherwise, if you are not interested sharing your results, i.e. in the above mentioned testing discount, please let me know and I could arrange for a discount when you place the order. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Thank you for your phone call yesterday and for your questions about ab6563. I have been in touch with the lab, and I was told that small amounts of precipitate are common and should not affect the viability of the antibody. It is possible that the antibody was damaged if it went through a few freeze-thaw cycles, but it should be fine stored at 4C for couple weeks. If you