Anti-Apolipoprotein B antibody (ab20737)

Below is information regarding the use of ab20737 in western blot.

1. Working dilution for immunoblotting - start low 1:100 - 1:500.
2. Molecular Weight of Apo B100 is 500kDa; Apo B48 is 250kDa. The western blot was run against purified LDL from mouse and against human plasma. Also tested mouse VLDL to detect ApoB48. When testing whole plasma, you may see additional bands (possible proteolytic fragments of ApoB100 and B48).
3. Use 1% BSA for blocking in the immunoblot part of the procedure.

Our general recommendations for transfer of large proteins, and a recommended paper that outlines a protocol for large protein transfer is given in our western blot guide below. As I mentioned, I recommended ensuring there is no methanol in your transfer buffer and including 0.1% SDS in the transfer buffer as well.

https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf

Below is a link to a paper that used ab20737 in western blot with mouse samples.

http://www.ncbi.nlm.nih.gov/pubmed/17438339?dopt=Abstract

Because we carry over 100,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If a species or application is untested you can use our products without financial risk by participating in our AbTrial program.

You would have to buy the ab117317 but once you submitted the Abreview you will receive a credit over the whole value of ab117317. You will receive this for either a positive and negative result of the testing.

I hope this information is helpful. Please let me know if you like to except the AbTrial offer and I would be happy to issue the Trial Code for you. If you have further questions please do not hesitate to contact me.

Thank you for your inquiry.

I am happy to confirm that we offer an Anti-Apolipoprotein B antibody that is tested and guaranteed for ICC and WB in rat, human and mouse samples:

ab117317b (Anti-Apolipoprotein B antibody)

https://www.abcam.com/index.html?datasheet=117317 (or use the following: https://www.abcam.com/index.html?datasheet=117317).

ab117317 is a chicken IgY antibody a suitbale secondary antibody is anti chicken IgY.

For example:

ab97135

https://www.abcam.com/index.html?datasheet=97135 (or use the following: https://www.abcam.com/index.html?datasheet=97135).

Apolipoprotein B is a very big protein (over 500kDa plus many modifications) and I would suggest to review these tips for big proteins in WB.

The following reference discusses a gel and buffer system that allows transfer of proteins as large as 500 kD:

Bolt and Mahoney, High-efficiency blotting of proteins of diverse sizes following sodium dodecyl sulfate–polyacrylamide, gel electrophoresis. Analytical Biochemistry 247, 185–192 (1997).

https://www.abcam.com/index.html?pageconfig=resource&rid=11404#C2

I hope this information is helpful and wish you good luck with your experiments.

Thank you for contacting us. Your credit note ID is xxxxx. I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Thank you for your reply.

I can gladly create this credit note/refund for you provided that you have purchase this item within the last six months. In order for me to issue this could you please provide me the Abcam order confirmation number for purchase of ab20737.

I look forward to your reply. Please let me know if you have any other questions.

Thank you for your reply. If your Abcam order was placed by credit card, the credit note indicates the dollar amount that will be refunded to your credit card account. Please allow 2-5 business days for the credit to process and appear on your statement. If your Abcam order was placed by Purchase Order andthe original invoice has not yet been paid to Abcam, your credit note will be forwarded on to your institution’s accounts payable department. They will subtract the credit amount from the invoice total, and disburse a check (if any) for the correct amount payable to Abcam. If your Abcam order was placed by Purchase Order andthe original invoice has already been paid, your credit note will be sent to your accounts payable department, and is credited back against the PO used for the purchase and put back into your budget. If your budget has been closed and this is no longer possible, you can request a refund by check instead at us.credits@abcam.com. The credit can also serve as a reference for a discount on a future purchase. I hope you have found this information helpful. Please do not hesitate to contact us with any additional questions you may have.

Thank you for taking time to provide this information and for contacting us. I am sorry to hear this antibody is not providing satisfactory results and that these suggestions did not help.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I wish to apologize for the inconvenience. If you have purchase this item within the last six months I am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Thank you for contacting us.

The expected molecular weight of this protein is 549KDa. As recommended by our https://www.abcam.com/index.html?pageconfig=resource&rid=11403#B1 the bigger the size of the protein of interest, the lower the percentage of gel. I would suggest using at least an 8% gel (as used by another user in the https://www.abcam.com/index.html?datasheet=20737&tab=abreviews&intabreviewid=12715 which gave excellent results) and using reduced and denaturing conditions.

I hope this helps. Please do not hesitate to contact us for further assistance.