Anti-Cytokeratin 18 antibody [C-04] (ab668)

HI,

I BOUGHT

Mouse IgG1 [ICIGG1] - Isotype Control (ab91353)

100 µg at 0.1mg/ml

2µg/1x106 cells


Anti-Cytokeratin 18 antibody [C-04] (ab668)

100 µg at 1mg/ml

1µg/1x106 cells

WHEN I USED THEM FOR FLOW I SAW THAT NEGATIVE WAS STRONGER AND HIGHER THAN POSITIVE.

DO I GET IT RIGHT THAT 1µg CYT18 - IS 1 MKL AND THE SAME FOR CONTROL ABS IS 10 MKL?

PLEASE HELP ME TO UNDERSTAND!!!

ALSO I USED PARAFORMALDEHIDE INSTEAD OF METHANOL AS IT SASY THAT STAINING WAS THE SAME.

AS I TRIED THIS FOR CELLS THAT WERE FIRST STAINED FOR SURFACE MARKERS I WAS AFRAID OF LOOSING SIGNAL AFTER METHANOL.

ANSWER:

Thank you for contacting Abcam.

I am sorry that you are having some issues using ab668 in flow cytometry. The antibody is covered under our Abpromise for six months and is guaranteed to work in flow cytometry on host of samples. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

To be able to aid you further, would you be able to send me a copy of the entire protocol that you use, including whether you permeabilized the cells or not and if you can send an example of the results you are getting, that would also be helpful.

Looking forward to your reply and helping you resolve this issue.

We are using ab668, mAb against cytokeratin 18, clone C-04. Is the epitope for this antibody known?

ANSWER:

Thank you for contacting us.

Ab668 is created by immunizing mouse with a Cytoskeleton preparation of epidermal carcinoma cell line A431. To our knowledge the exact epitope for this antibody has not yet been mapped.

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

Customer is using this antibody in WB with human liver cells. At dilutions of 1:1000 and 1:5000, is seeing a clean doublet at approx 45 kDa.

Has this been reported before with this antibody? What is recommended as a positive control for WB?

ANSWER:

Thank you for your enquiry.

This protein can be glycosylated and has various variants so I suspect that you are seeing two forms of the protein, i.e glycosylated isoforms or partially degraded CK18 (by natural cleaved mechanisms or during sample preparation).

We use the cell line A431 for testing ab668 but any line expressing CK18 works well as this clone is one of the best CK18 markers.

I hope this information will help you, please let me know if you need anything else.

I would like to pick ab9287.

By the way, when viewing the list of cytokeratin abs, there seems to be other products named as "cytokeratin 8" antibodies but reacts specifically with cytokeratin 18. For examples, ab7797/ab7798.

ANSWER:

For ab7797 and ab7798, you're right! I have changed those datasheets as well.

Ab9287 is being sent to you free of charge on order# 72784 and you should receive it today or tomorrow.

Please let me know if you have any additional questions.

i have an old vial of this antibody (ab668-100) that says "mouse monoclonal (c-04) to cytokeratin 18. however when i went to your website and looked at this catalog number, it says cytokeratin 8, instead of 18. would you please clarify this to me. also, there is the cytokeratin 8 antibody sc9023. i'd like to know which one is keratin 8 or 18 and whether anyone of them recognises both proteins or are specific to one of them. thanks

ANSWER:

Thank you for your message.

Ab668 is a Cytokeratin 18 antibody, NOT a Cytokeratin 8 antibody. There was a mistake on the online datasheet which we have now updated.

I'm not sure what you mean by "cytokeratin 8 antibody sc9023." Do you have the Abcam catalog number?