Anti-iNOS antibody (ab3523)

We have verified the antibody ab3523 twice again in our lab, it appears the antibody does not perform with sufficient dilution factors. No matter how high we raised the dilutions it still appears negative. For instance at dilution 1:10.
We have to conclude that this antibody does not appear optimal performance on dilutions. We thereas ask for refund of this antibody.
Thanks for your time and review.
Thanks a lot

ANSWER:

Thank you for contacting us.

Your credit note ID is XXXXXXX.

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.

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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.

Crystal from biomole

ANSWER:

Thank you for contacting us.

Please be advised that we can not provide refund or free of charge replacement because Bimole is not an official distributor of Abcam. Our policy is that we will offer pre- and post-sale technical support for products sold via unauthorized distributors. We will not offer our guarantee as we cannot control the shipping and storage conditions managed by the unauthorized distributor.

We unfortunately have no further recommendation for this product, we suggest customer to buy the antibody form Abcam website.

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

Please find the attached as instructed. Look forward to your suggest.
Thanks

ANSWER:

Thank you for your email.

After reviewing the details again I would suggest using RIPA lysis buffer, which gives best results and using fresh tissues for lysates preparation.

I am sorry we do not have any further recommendation to make. The antibody should be working with the conditions you have used however we are unsure why the antibody failed. Please also be advised that we can not provide refund or free of charge replacement because Bimole is not an official distributor of Abcam. Our policy is that we will offer pre- and post-sale technical support for products sold via unauthorized distributors. In case of problems with our antibodies we will offer suggestions to the researcher's protocol but we will not offer our guarantee as we cannot control the shipping and storage conditions managed by the unauthorized distributor.

I hope this informaiton is nevertheless helpful. SHould you have any other question please do not hesitate to cotnact me.

Thank you for your suggestion! After blocking with 5%BSA in PBS, should I
also probe the membrane in the same 5%BSA in PBS?

ANSWER:

Thank you for your reply.

Yes, along with using the 5%BSA to block the membrane, I would also suggest using the 5%BSA to dilute the primary and secondary antibody in as well.

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

I bought the anti-iNOS antibody (ab3523). On my western image, there seem
3 bands above 120 kd, we are not sure which band is iNOS, please give
advice.
The samples are mouse liver tissue lysate, 50ug protein, run on 12% gel,
tranferred to nitrocellulose membrane. Block in 5% milk TBST for 1 hr.
Anti-iNOS 1/200 in 5% milk TBST in the fridge overnight. Wash with TBST3X5 min. Goat anti-rabbit HRP (Thermo Sci) 1/5000 in 5% milk TBST at RT
for 1 hr. Wash 3X5 min. Add the substrate (SuperSignal, Thermo Sci) for 5
min. The membrane is exposed in Fujifilm LAS-4000 for 20 sec. The top 3
bands of marker are 220, 120,100 kd.
Attached are original image file and the image in ppt.

ANSWER:

Thank you for sending that information.

I have placed red arrows next to the bands that I think are the iNOS signal. When you redo the western blot, I would recommend using 5%BSA in PBS as your blocking reagent, as this will help to remove some of the background bands you are seeing. Thereby allowing you to distinguuish your band of interest clearer.

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