Anti-CD63 抗体 [MEM-259] (ab8219)

ab2215 should be used at a concentration of 2-4 ug/ml in blocking buffer when incubating your membrane whereas ab8216 should be used at a concentration of 2 ug/ml.

The gel should be run in non-reducing conditions as these antibodies do not recognise their antigens well when the western blot sample contains reducing agent. We therefore advise against adding beta-mercaptoethanol or DTT in the loading buffer.

We have not tested the stability of ab8219 under the alkaline conditions that you described; thus, we cannot guarantee that it will work under these conditions. The laboratory thinks it is possible that it could work, but only performing the experiment will tell.

We are not aware of a reference describing production and characterization of this CD63 antibody. The epitope the antibody recognizes on CD63 has not, to our knowledge, been mapped, but we do know that the epitope is extracellular when CD63 is integrated into the plasma membrane.

Thank you for contacting us.

The antibodies have not been tested for reactivity with bovine samples, and we have not received reports of bovine reactivity from our customers.

Please do not hesitate to contact us if you need any more advice or information.

Thank you for your inquiry.

Ab8219 (anti-CD63) is supplied as 100ul at a concentration of 1mg/ml. We have a customer who reported successfully using the antibody at a 1/200 dilution (0.2 mg/ml), but it may be necessary to optimize this for your own experiments. Please note that we recommend using non-reducing conditions for the western blot (i.e. no BME or DTT).

I hope this information helps. Please do not hesitate to contact us if you need anything further. Have a wonderful day, and best of luck in your research!

Thank you for your enquiry.

I can confirm that the epitope for ab8219 CD63 antibody [MEM-259] has been mapped to the extracellular region of the CD63 protein.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Thnak you for contacting Abcam.

In regards to ab8219, I have had the chance to talk with the lab. According to them this antigen is indeed very variable on WB of various material. I am sending the pattern they obtained on non-reducing K562 cells (see image CCE3541736_MEM-259WB.jpg), however observing more heavier bands using other material would be also a reasonable result.

I have attached information given to me by the lab producing ab59479 (see remainng attached). These include a puplication which has used the product in Western blot as well as a Powerpoint file which contains lab testing results.

I hope this information is helpful. Please do not hestitate to contact me if you have any questions.

Thank you for contacting us.

CD63 antigen is also known as LIMP and LAMP-3. While unmodified protein is about 25kDA, this is heavily glycolylated. AB59476migrates like a smear due to a very complexe N-glycosylation. I would recommend this product because this real world information is available whereas the data for ab8219 appears to be based on theory and not practise.

Please remember that differences exist between the cell lines (e.g. K562 givesbetter migration than HEL).

It is important to remember that this antibody (as well that all theanti-tetraspanine antibodies) must be used under non-reduced conditions.



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

The antibody was validated for western blotting using these conditions:

Sample: K562 cells (24 millions, lysed with 1% laurylmaltoside in 700 ul), non-reducing SDS, denatured by heat for 5 min.
Blocking: 5% milk, 4oC overnight
Primary antibody: diluted 1:100 in blocking solution, room temperature, 60 min.

I hope this is helpful. If you have any questions, please do not hesitate to contact us.