Anti-Arp5 antibody (ab12099)

it seems to me looking on your Western Blot that a-Arp5 antibody, cross interacts with Arp4 (band at 70 kDa). For my application it is essential that there is no cross-reactivity to Arp8 or Arp4.

I have the similiar question concerning your Arp8 antibody. In the WB there are also additional bands (not the degradation).

Do you know if on the WB of Arp5 of the knock-out studies the additional cross-reactive bands (not shown) vanish?

Would it be possible to send me small aliqouts of those antibodies, that I can check the cross-reactivity?

As like I said it is essential for our us to know this, before we order the antibodies.

ANSWER:

Thank you for contacting us.

I have received the following information from the lab:

Ab12099 (ARP5):

Unfortunately, the full WB image showing the knock-out experiment is not on file, thus we are not able to determine if there were additional bands on the blot. We believe the testing may have been done by a collaborator a while ago, but currently do not have information to confirm that.

We have not tested for cross reactivity with Arp4 or Arp8. The immunogen is the full length recombinant protein, thus we do not know where the epitope is located.

The sequence homologies for the Arp5 yeast protein are as follows:

Arp5-Arp4 = 21-30% (with lots of gaps)

Arp5-Arp8 = very poor (30-53% for very short stretches, have lots of gaps)

Thus, crossreactivity is quite unlikely. Usually a homology of 85% or higher is likely for cross reactivity, but not as low as 30%.

Ab12098 (ARP8):

We have no information on whether this antibody crossreacts with Arp4 and Arp5. The immunogen for this antibody is the full length recombinant protein. Again, we do not know the epitope region as it has not been mapped.

The sequence homologies for the Arp5 yeast protein are as follows:

Arp8-Arp4 = very poor (22-38% for short stretches, have lots of gaps)

Arp8-Arp5 = very poor (30-53% for short stretches, have lots of gaps)

Again, crossreactivity is quite unlikely as stated above.

The expected MW of the unprocessed precursors for each yeast protein are

Arp5 = 87 kDa

Arp8 = 100 kDa

Arp4 = 55 kDa

Any other bands of different MW could be non-specific bands (but not Arp4,5 or 8).

Unfortunately, we do not send out small aliquots as this is operationally not feasible for us. However, we do guarantee all our products for tested species and applications as listed on the datasheet. The guarantee is valid for 6 months after a purchase. For more details about our Abpromise guarantee please check this link: http://www.abcam.com/abpromise.

Should the antibodies not give you satisfactory results, we will be able to replace, credit or refund them.

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