Anti-P Glycoprotein antibody [JSB-1] (ab3366)

Thank you for contacting us. The concentration of that product is 67ug/ml. If you are looking to concentrate an antibody, we recommend using our Antibody Purification kit (102783). At Abcam, providing quality service is just as important as providing quality products. Our Abpromise® ensures that you can trust our products to perform as stated on the datasheets, or we will offer a replacement, credit, or refund.   Another option that may work for you, because this is a high purified product, would be to use protein G beads to capture the antibody and re-suspend in a lesser amount of buffer. I cannot guarentee this but thought you might want to look into that option as well.  I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

Thank you for contacting us in regard to this issue. I've looked into your question and have found that the concentration is lot specific. A recent lot was 167 ug/ml. If you have ordered already, could you give me the lot number of the vial that you have? Please do not hesitate to contact us with further questions or if there are other ways in which we might help you reach your research goals.

Vielen Dank für Ihre Anfrage und dafür, dass Sie sich die Zeit genommen haben, uns das Problem so detailiert zu beschreiben. Es tut mir leid, dass Sie Probleme mit unserem Antikörper haben. Ich habe noch ein paar Fragen zu Ihrem Protokol: 1) Was ist die Spezie Ihrer Proben und der positiven Kontrolle (human?)? 2) Welche Bestandteile hat der Lysispuffer und welches Detergent ist enthalten? Das Protein is ein hydrophobes Protein mit 12 transmembranen Domainen. Die Probenherstellung ist daher ein wichtiger Schritt, ebenso wie der Proteintransfer vom Gel zur Membran. Es is wichtig, dass das Protein genuegend solubilisiert wird damit es in Loesung bleibt und auch beim Transfer nicht im Gel prezipitiert. Von dem WB den Sie mir geschickt haben, sieht es so aus, als ob das Protein nicht detektiert wurde, weil moeglicherweise nicht genuegend Protein (d.h. Glycoprotein P) auf der Membran ist. Das Protokol, dass Sie benutzen, ist meiner Meinung nach in Ordnung. Hier sind ein paar Tips, die bei der Probenherstellung und Proteindetektion hoffentlich weiterhelfen werden: 1) Ich bin mit der Zusammensetzung des CLB1 Puffers nicht vertraut, wuerde aber vorschlagen den RIPA buffer anstatt zu verwenden. Der RIPA buffer enthaelt 3 Detergenzien: SDS, NP-40 und Natrium deoxycholate, die bei der Solubilisierung vom Membranproteinen sehr hilfreich sind. Hier ist der Link zu unserer Webseite "Puffer und Stockloesungen": https://www.abcam.com/index.html?pageconfig=resource&rid=11414 RIPA buffer (radio immuno precipitation assay) buffer: RIPA buffer contains the ionic detergent sodium deoxycholate as an active constituent and is particularly used for nuclear membrane disruption for nuclear extracts. A RIPA buffer gives low background but can denature kinases. It can also disrupt protein-protein interactions (and may therefore be problematic for immunoprecipitations/pull down assays). 50 mM Tris HCl pH 8 150 mM NaCl 1% NP-40 0.5% sodium deoxycholate 0.1% SDS The 10% sodium deoxycholate stock solution (5 g into 50 ml) must be protected from light. The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O and then add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the total volume to 50 ml). Store the buffer at 4°C. 2) Ich wuerde ebenfalls raten (falls Sie das nicht schon machen) dem Transferbuffer bis zu 0.1% SDS hinzuzufuegen. Das hilft eine Prezipitation von hydrophoben Proteinen zu vermeiden. 3) Ausserdem waere es moeglich, die Effektivitaet des Transfers zu ueberpruefen, entweder mit einer CoomassieBlue Faerbung des Gels (nach dem Transfer) und/oder einer Ponceau S Faerbung der Membran. Im erwarteten Bereich von 140 kDa sollten Sie ein Signal sehen mit Ponceau S, waehrend die Coomassie Faerbung des Gels sollte Ihnen im Idealfall keine Anfaerbung zeigen. Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte und zögern Sie nicht, sich wieder bei uns zu melden, falls dies Tips Ihnen nicht geholfen haben.

The concentration of lot # GR46599-2 is 66.7ug/ml. If you have any other questions please feel free to contact us.

Thank you for contacting Abcam regarding ab3366 I have confirmed with the laboratory that lot 963386 was made at 161ug/ml, which is probably why you are seeing a difference. There is lot to lot variation in concentration, however, staining at 1:40 should be consistent. I would recommend testing this dilution to see if your results improve. I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions or if this suggestion does not improve your results with this antibody.

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.

Thank you for your telephone enquiry yesterday. I can confirm that the concentration of ab3366 is 100ug/ml (there may be slight variation from lot to lot). I hope this information is helpful. Should you have any further questions, please do not hesitate to contact us again.

Thank you for your enquiry. I am sorry it has taken a very long time to obtain details from the laboratory for you. However, the concentration for the particular lot number of the antibody you are using (lot 208647 ab3366) is 80ug/ml. The concentration you use it at will depend on optimisation for your own samples. I have copied a link here to our IP protocol web page which may help you decide: http://ops.abcam.com/index.html?pageconfig=resource&rid=10416 I hope this helps. Once again, thank you for your patience. If there is anything else you need to ask, please get in touch again and I will be happy to help. With thanks

Thank you for the images. Does your sentence "Besides the same second antibody can be work in another IHC detection with Cat#ab3366" mean that the secondary antibody works well? The colon tissue must be colon carcinoma, not normal colon tissue as the latter does not have high levels of MLH1. Please provide a detailed protocol of the customer's staining (using the Abcam questionnaire pasted in the body of an e-mail or via our website). In the future please provide this information as soon as you contact us so we minimise the delay and inconvenience for the customer, Thank you for your support,

Thank you for your enquiry. ab3366 (JSB-1) was tested using HIER (heat-induced epitope retrieval) at a high pH (EDTA, pH 8-9). I apologize for the datasheet not containing this information. Please re-try the experiment using these conditions and contact us if you do not get a positive result. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.