Anti-PAG antibody (ab14989)

I would actually like to carry out both Western blotting and IP. So referring to the datasheet, what step would you recommend to add 1% laurylmaltoside. And is this to subsitute other degerents used in the assay? The detergent that I use in my SDS-PAGE is SDS and B mercaptoethanol and I perform a boiling before loading onto the gel. The detergent that I use for Western blotting is Tween-20.

Could you maybe send me the detailed procedure/protocol for WB and IP? Our lab is currently working on Cbp in different systems and it will be great if we can find a good antibody for it.

ANSWER:

My apologies for the late reply, I would like to help you as my colleague Tanya is away:

Laurylmaltoside is a detergent for cell lysis buffer not for SDS-sample buffer. I have just been informed by the supplier that the antibody should stain also reduced PAG antigen. Detailed protocols have been published by the V. Horejsi group (founders of Csk-binding protein), Horejsi's paper published in J.Exp. Med. is available free online. Please try: - Brdicka T. et al. J. Exp. Med. 191, 1591 (2000). - Brdickova N. et al. FEBS Lett. 507, 133 (2001).

I hope this information helps, please do not hesitate to contact us again if you need further advise,

I would actually like to carry out both Western blotting and IP. So referring to the datasheet, what step would you recommend to add 1% laurylmaltoside. And is this to subsitute other degerents used in the assay? The detergent that I use in my SDS-PAGE is SDS and B mercaptoethanol and I perform a boiling before loading onto the gel. The detergent that I use for Western blotting is Tween-20.

Could you maybe send me the detailed procedure/protocol for WB and IP? Our lab is currently working on Cbp in different systems and it will be great if we can find a good antibody for it.

ANSWER:

My apologies for the late reply, I would like to help you as my colleague Tanya is away:

Laurylmaltoside is a detergent for cell lysis buffer not for SDS-sample buffer. I have just been informed by the supplier that the antibody should stain also reduced PAG antigen. Detailed protocols have been published by the V. Horejsi group (founders of Csk-binding protein), Horejsi's paper published in J.Exp. Med. is available free online. Please try: - Brdicka T. et al. J. Exp. Med. 191, 1591 (2000). - Brdickova N. et al. FEBS Lett. 507, 133 (2001).

I hope this information helps, please do not hesitate to contact us again if you need further advise,

Hello!

Please let me know this antibody #ab14989.

The data sheet of this antibody has a publication as WB in a mouse is possible. Do you have the data of actually having been used for WB of a mouse, in your company? Or when you have the paper used such, please introduce. In HP of your company, it was displayed that there was no used paper at present.

Thank you for your cooperation.

ANSWER:

Thank you for your enquiry.

We would like to confirm that this antibody has been tested and characterised for Western blot analysis and it recognizes mouse PAG.

Unfortunately, there is no published reference available yet. We will update the on-line datasheet of this product as soon as get more data.