Anti-P Glycoprotein antibody [4E3] (ab10333)

ab10333 gives no band in WB on human cancer cells

ANSWER:

Thank you for your phone call.
As we discussed, ab10333 has not been validated for use in WB and may or may not be suitable for use in that application. While it will not be covered under our Abpromise guarantee, the following may help to improve your results:
1) Because P Glycoprotein is a multi-pass membrane protein, it will tend to aggregate when boiled. To avoid this, I would recommend heating to a lower temperature (around 70C) for 10-20 minutes to achieve denaturation.
2) P Glycoprotein is also a high molecular weight protein. To run effectively, the percentage of the gel will need to be 8% or lower.
3) High molecular weight proteins may not transfer properly using standard buffers. Adding 0.1% SDS to the transfer buffer and keeping the methanol concentration to 10% or lower will help the transfer efficiency. It is recommended to validate the transfer with a ponceau stain to ensure the larger proteins have transferred properly.
4) In general, you may be able to increase your signal by using more primary antibody or incubating overnight at 4C.
I hope this helps, if not, please provide a brief outline of your protocol information and I will be happy to assist you further.

The epitope information for this antibody says, "The 4E3 monoclonal antibody recognizes a cell surface epitope of the 170kDa multidrug resistance P-glycoprotein." Which extracellular epitope is it?

ANSWER:

Thank you for your inquiry.


I heard back from the supplying lab that theyhave not mapped the exact epitope for this antibody. We have licensed this from the inventor of the antibody. The only information we have about it was provided by Dr Arceci's lab. The following publication describes the invention of this antibody, however it doesn't have a lot of information regarding epitope.


Arceci RJ, et al. Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein. Cancer Res. 53:310-317, 1993



I'm sorry I cannot provide more insight into this. I did a quick literature search and it doesn't appear that anyone has published information regarding the epitope of 4E3, so unfortunately we do not know which extracellular epitope it binds. I apologize that we have not been able to help you in this regard, but please let me know if you have any other questions.

Dear Sir/Madam,

I was recently reading an article which uses one of your antibodies. I cannot seem to locate this product on your website and was wondering if you could point me in the right direction. The article states using:

"a mouse-monoclonal antibody against mammalian Pgp (C219; Abcam, Cambridge, MA, USA).... This antibody targets two Pgp epitopes that are largely conserved in the SMDR2 sequence"

Another issue you may be able to help me with is whether or not you have a primary antibody that would recognise a variety of pgp/mdr proteins (has to include MDR2 (human gene family ABCB1) and MRP1 (human gene family ABCC1)) i.e. it may bind a highly conserved region across the pgps.

I would really appreciate any help with the matter as I have been trawling through many of the antibodies available and comparing immunogens.

I hope you can help, thank you for your time

Kind regards

ANSWER:

Thank you for contacting us.

ab3364; anti P Glycoprotein is no longer available in our catalogue. However we have similar products available to buy. The catalogue numbers are

ab3083; ab3366; ab10333; ab80594; ab80626. Please check the datasheets of these antibodies and select a product according to your requirements.

The anti MDR2 products are ab71792; ab24108 and ab111209

Anti MRP1 products are ab24102; ab63987; ab99531; ab81903

To search different products please type target name in the search box provided on the Abcam homepage at www.abcam.com. After hitting the search button the system will give you the sorted product as per your target request.

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

Did you receive the following email I sento to you? I haven't received your confirmation.

>Thanks for your reply. >I have contacted with the customer, she prefer you to give a credit, so please issue one about this item for us.

ANSWER:

Thank you for your email. I do apologize as I think that a reply got lost somehow in the system.

For this antibody, we were selling it in a smaller size (100 ul) but it did not provide enough tests. We are now providing it in a 500 ul size. If your customer is interested, I can send you the 500 ul vial free of charge, or I can provide you with a credit. Please let me know which is preferred.

One customer have bought one vial ab10333/MDR1 antibody [4E3] from you. the volume is 100ul and the concertration is 0.1mg/ml, so the total quantity is 10ug. the size is too small and the recommendated dilution is not clear on the datasheet. it's difficult for the customer to use it. So can you give us the recommendated dilution for application indicated on datasheet.

ANSWER:

Thank you for your enquiry.

We have received the following information from the source of ab0333:

"We recommend that the customer start with a 1:10 dilution and titer Ab from there for their individual application."

In light of this information we can certainly offer you a refund or credit note and we are considering removing the antibody from our catalogue, I am sorry about this inconvenience,

Please let me know if your customer wishes a credit note and we will issue it to you,