Anti-EID1 antibody (ab14454)

BATCH NUMBER Lot79677

DESCRIPTION OF THE PROBLEM No signal

SAMPLE We tested its reactivity towards the endogenous EID1 in 100microgram lysates from HeLa, U937 and 293 cells, and also on 293 cells over-expressing high levels of exogenous EID1.

PRIMARY ANTIBODY 4 microgram/ ml, 1h or overnight

DETECTION METHOD ECL-Pierce's super-sensitive ECL reagent.

POSITIVE AND NEGATIVE CONTROLS USED antibody to this protein from an alternate source worked just fine under same conditions.

ANTIBODY STORAGE CONDITIONS 4c

SAMPLE PREPARATION RIPA +protease inhibitors, boiling 5 min with protein sample buffer that contains beta mercaptoethanol

AMOUNT OF PROTEIN LOADED 100 microgram total protein

ELECTROPHORESIS/GEL CONDITIONS 10%, 12% SDS PAGE Gel

TRANSFER AND BLOCKING CONDITIONS 1.5 h transfer Tris glycin buffer, blocking with 10% milk

SECONDARY ANTIBODY We used horseradish peroxidase linked-protein A (Amersham) as our secondary. Pierce's super-sensitive ECL reagent.

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

ADDITIONAL NOTES We used this product for western blots, according to your recommendations in the product datasheet, at a concentration of 4microgram/ml (i.e. the higher concentration). We tested its reactivity towards the endogenous EID1 in 100microgram lysates from HeLa, U937 and 293 cells, and also on 293 cells over-expressing high levels of exogenous EID1. We used horseradish peroxidase linked-protein A (Amersham) as our secondary, and Pierce's super-sensitive ECL reagent. However, we failed to detect any signal with your antibody, although a second antibody to this protein from an alternate source worked just fine. As this product does not seem to work, I would like to be able to return it and get a refund.

Thank you for yor assistance,

ANSWER:

Thank you for your enquiry and for taking your time to fill in the on-line Questionnaire and for providing details of your Western blot protocol. I am very sorry to hear that you are having problem with this antibody.

I have looked through your procedure and it looks absolutely fine. I have also checked FedEx website and the delivery details in our computer system and found the following. The package was held up at the customs for clearance and checking and it may well be that during this time the antibody was not stored properly.

Since you have an alternative antibody which works nicely, certainly I can offer you reimbursement in this case. I can offer you either a credit note which you can use for your next purchase or a full refund.

Please do let me know how you would prefer to proceed and also confirm your Purchase Order Number and/or your Abcam Order Number.

I am looking forward to hearing from you soon.

wanted to double check that the EID1 antibody (ab14454) cross react with mouse EID1

ANSWER:

Thank you for your enquiry regarding ab14454.

We have not yet tested the cross-reactivity for this antibody with mouse tissue but expect the antibody to work as there is 90% similarity between mouse and human (which we have tested).

Should you decide to test an antibody in mouse please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.