Anti-Pma1抗体[40B7] (ab4645)

Thank you very much for your reply.

The anti-IgG (H&L) secondary will bind to the conserved light chains of the IgM primary. I suggest using the secondary at a higher concentration than you normally would with an IgG primary, but I'd expect this secondary to be fine. You can try increasing the primary to 1:500, and the secondary to 1:5000 to see if this helps.

To confirm solubility of membrane proteins you could try probing for another membrane marker, if you have an antibody to a different target available. I'm not sure of a better way to confirm this, but if the proteins didn't fully extract then this could be the cause of no signal. We do have one other antibody for an S. cerevisiae membrane protein, ab110326-

https://www.abcam.com/VDACPorin-antibody-16G9E6BC4-Mitochondrial-Loading-Control-ab110326.html

Would this be helpful for you? I look forward to hearing from you after discussing with your PI. Please let me know if you have any questions or if there is anything else that we can do for you. Have a nice weekend!

Thank you very much for contacting us and letting us know about the trouble with this antibody.

Thank you also for sending so many details about your protocol. I do have a couple of questions, just so I can further understand the situation:

1) Was the transfer to PVDF verified with a Ponceau stain or another method?
2) Were the secondary antibodies anti-mouse IgG or anti-mouse IgM?
3) What kind of samples you using and what species are they from?

I would expect the antibody to work under the conditions you've described, but it may be helpful to load more protein and increase the concentration of antibody. I understand that you have already tried some troubleshooting, and this antibody is guaranteed to work in WB with S. cerevisiae samples, so if you would like a replacement, credit, or refund, I'll be happy to set that up for you.

I look forward to hearing from you. If you have any further questions or need anything else, please let me know and I'll be happy to help.

Thank you for contacting Abcam.

Under our Abpromise, we guarantee this antibody to work in western on yeast samples. As we had not tested in oh human samples, then we could not guarantee that it would work on this species.

I would be very happy to help you troubleshoot though and see if we can try and get the antibody working. A few protocol variations that may help include:

1 - Block in BSA, as this would help to reduce background and allow a longer exposure to see the 100kDa bands, if the antibody detects human samples.

2 - Increase the total amount of protein loaded to 40ug.

3 - Increase the primary antibody concentration from 1/7000, to 1/1000.

If the above protocol variations do not prove to be successful, then please let me know and we can try and come up with other solutions.

Also for your future reference, if you would ever like to use on of our antibodies in an untested application or species, then you could take advantage of our testing discount program:

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount



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

Thank you for your enquiry.

Below is a selection of antibodies that are suitable for detecting the plasma membrane and nuclear envelope. They all work in western blot and I would review the datasheets to confirm which species they are tested against.

Plasma membrane
ab4645 Anti-Pma1 antibody [40B7]
www.abcam.com/4645

ab7671 Anti-alpha 1 Sodium Potassium ATPase antibody [464.6]
www.abcam.com/ab7671

ab16505 Anti-pan Cadherin antibody
www.abcam.com/ab16505

ab22744 Anti-pan Cadherin antibody [mAbcam22744]
www.abcam.com/ab22744

Nuclear envelope
ab8980 Anti-Lamin A antibody [133A2]
www.abcam.com/ab8980

ab8984 Anti-Lamin A + C antibody [131C3]
www.abcam.com/ab8984

ab16048 Anti-Lamin B1 antibody
www.abcam.com/ab16048

I hope this is useful. Just let me know if you have further questions.

Merci beaucoup pour votre intérêt pour nos anticorps. Nous avons trois anticorps actuellement qui pourront correspondre a vos besoin. Veuillez vérifier leur fiche technique s'il vous plaît. Je n'ai pas trouvé un anticorps testé contre pichia pastoris par contre. Dans quels applications est-ce que vous avez besoin d'utiliser cet anticorps? Anti-PDI antibody [PDI-38H8] - ER Marker (ab30319) Click here (or use the following: https://www.abcam.com/P4HB-antibody-PDI-38H8-ab30319.html). ti-Ribophorin I antibody (ab52672) Click here (or use the following: https://www.abcam.com/Ribophorin-I-antibody-ab52672.html). Anti-KDEL antibody [10C3] (ab94746) Click here (or use the following: https://www.abcam.com/HUS1B-antibody-ab4746.html). J'espère que ces informations vous sont utiles et que un des anticorps ci dessus peux correspondre à vos besoins. N'hésitez pas à nous recontacter si vous avez des autres questions.

Thank you for your enquiry. The isotype of this antibody is IgG3 with kappa light chains. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soon as any new information is brought to our attention. As far as we are aware, cross reactivity with Canida albican has not yet been tested for use with ab4645. Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher. Please let me know if you require any further assistance.

The Pma1p protein is very antigenic, and antibodies to it turn up frequently if yeast membrane preps are used as the immunogen. However, what the exact antigenic region is, is not currently known.