Anti-P180 Lamellar Body Protein antibody [3C9] (ab24751)

When using this antibody for FACS, will the cells require permeabilisation? Is the epitope on the intracellular section of the membrane?

ANSWER:

Thank you for your telephone enquiry yesterday.

Unfortunately, the epitope for this antibody (ab24751) has not been mapped, so it is not known if it is on the outer membrane or on the intracellular section of the protein.

I can suggest trying permeabilisation. This should be with a more gentle reagent. Tween 20, Saponin, Digitonin and Leucoperm are mild membrane solubilisers. Use at 0.5% in PBS. These give large enough pores for antibodies to go through without dissolving plasma membrane Harsh detergents such as triton or acetone are not suitable for membrane proteins.

We are always pleased to hear from customers who have used antibodies in untested applications. If you decide to go ahead and purchase this product, we would be pleased if you can consider submitting some feedback by submitting an Abreview via the online product datasheet. We always encourage customers to send their results back to us, whether positive or negative, and we make all product information available to researchers.

To find out more about our Abreview system, please see the following URL: http://www.abcam.com/abreviews

I hope this information is helpful. Should you have any further questions, please do not hesitate to contact us again.

How was this antibody used in Western blot? What percentage gel was run? Was it reducing or non-reducing?

ANSWER:

Thank you for your enquiry.

The antibody was tested on a 5-15% gradient gel using reducing conditions.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Can you please tell to which portion (region) of the molecule this antibody has been generated? If you can give me the exact amino acid sequence for the immunogen, it would be very helpful.

ANSWER:

Thank you for your enquiry.

This antibody was raised against purified rat lamellar body membranes. We do not know the epitope, as epitope mapping was not done.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information,