Anti-Apolipoprotein E antibody (ab7620)

We have actually tested out the Apoe antibody on IHC-P in rat brain and it worked beautifully. I can post photos in a review on the website or submit them here if you would like, along with the protocol we used. From previous experience, I think we still get a credit for a free antibody for being the first to test an antibody in a certain application/species. Can we still use the code below?

Thank you!

ANSWER:

Thank you for your reply.


I am happy to hear that this antibody worked well for you in IHC-P. Please be advised that the code that was issued to you is a credit note, since the information on our website was found to be incorrect. It can be used at any time and towards any invoice.


This is separate from our testing discount option. For testing discounts, customers must contact us prior to purchasing a product to determine if a product is eligible for the discount. Once this is determined, a code is issued. In order to redeem the code, you must purchase the antibody you wish to test, test it in an untested application and/or species, and submit an Abreview of your results. Once the review is submitted, the code can be redeemed for a free primary antibody on a future order.


Please visit our website for additional details: http://www.abcam.com/collaborationdiscount.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

I was wondering if you had any photos from IHC-P of the Apoe antibody (ab7620).

ANSWER:

Thank you for your inquiry.


I heard back from the supplying laboratory that this antibody has not, in fact, been tested in IHC-P. Data for IHC-P for their anti-ApoA2 antibody (which is produced to similar specifications) suggests the anti-ApoE antibody will work well for IHC using suitable samples and conditions, however the ApoE antibody has not been validated in IHC-P. I have since removed this application from the list on the datasheet, and I sincerely apologize for the error.


Since ab7620 hasn'tbeen validated in IHC-P,I can offer a discount off a future purchase if you buy ab7620 now, test it in IHC-P and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary 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 ab7620 in IHC-P. I will then send a discount code. This code must be issued before purchasingab7620 so please wait for my reply before ordering.

2. Purchase ab7620 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-P.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: http://www.abcam.com/abreviews.

5. After the review 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 anyprimary antibodyordered 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 ab7620 turns out to be unsuitable for IHC-P, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

BATCH NUMBER 17739 ORDER NUMBER ab7620

DESCRIPTION OF THE PROBLEM wrong band size at 85 kDa

SAMPLE plasma derived enriched chylomicrons, ldl, hdl and plasma

PRIMARY ANTIBODY ApoE (Abcam #ab7620 as above) 1:5000 in PBS with 0.3% Tween with 2% Blotting Grade Blocker Non-Fat Dry Milk (Biorad)for 1h, three times washing with PBS with 0,3% Tween 20

SECONDARY ANTIBODY Donkey Anti-Goat IgG (H+L) (Jackson; Cat# 705-036-147)1:20000 in PBS with 0.3% Tween with 2% Blotting Grade Blocker Non-Fat Dry Milk (Biorad)for 1h, three times washing with PBS with 0,3% Tween 20

DETECTION METHOD ECL

POSITIVE AND NEGATIVE CONTROLS USED plasma derived enriched chylomicrons, ldl, hdl

ANTIBODY STORAGE CONDITIONS -20?C

SAMPLE PREPARATION precipitated proteins resolved in laemmli buffer (according to Jouranl of Lipi Research Volume 30, 1989 S757-761 (Maguire GF, Lee M, Connelly PW)Sodium dodecyl sulfate-glycerol polyacrylamide slab gel electrophoresis for the resolution of apolipoproteins.

AMOUNT OF PROTEIN LOADED 27 ?g protein

ELECTROPHORESIS/GEL CONDITIONS reducing (mercaptoethanol),12% SDS PAGE Gel

TRANSFER AND BLOCKING CONDITIONS 25mM Tris; 190mM Glycin; 1mM EDTA; 0.05% SDS; 20% Ethanol, time 1h at 60 V; 5% Blotting Grade Blocker Non-Fat Dry Milk (Biorad)in PBS with 0.3% Tween 20

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

WHAT STEPS HAVE YOU ALTERED? the first dilution of the primary antibody was 1:1000 which was too less diluted because we have multiple unspecific bands and with 1:10000 dilution the signal was to weak there was only a signal at 85 kDa

this unspefic band at 85 kDa we still have at a antibody dilution of 1:5000 which we used in the later experiments

ANSWER:

I'm sorry to hear you are having problem with ab7620. This is the first complaint we receive about this popular antibody.

We would recommend using this antibody at a 1:5000 to 1:10 000 dilution, and I am puzzled at why you detect an 85kDa band. Having researched on what possible reasons this might be, I have found out that the protein has several isoforms, a potential carbohydrate chain can be added onto some forms, and it has several binding sites: for heparin (5-30kDa), beta amyloid as well as the receptors VLDL (160kDa) and apoER2 (10.2kDa). I have contacted the source of this antibody to ask for suggestions and also for the protocol used to test the antibody. Unfortunately there may be a delay of more than a week as the person is currently away.

My sincere apologies for the delay,

ANTIBODY CODE ab7620

PRE-SALES ENQUIRY Does this antibody react with the E2 and E4 isoform of Apo E?

Thanks

ANSWER:

Thank you for your enquiry. The antibody was made against whole apo E. It should react with all isoforms which enrich in Human plasma. If you have any further questions please do not hesitate to get in touch.

I would like to purchase an antibody with immunoreactivity against mouse Apoliprotein E. Has your antibody been tested against samples from mouse cell line? If not, would it be possible for you to send a small test sample to the above address? or to test the immunoreactivity against mouse apoE in your laboratory?

ANSWER:

Thank you for your enquiry. Ab7620 has been tested with cross-reactivity with human and has not yet been tested in other species.

We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement.

Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.