Goat F(ab')2 polyclonal Secondary Antibody to Mouse IgG - H&L (PE), pre-adsorbed (ab7002)

thank you very much for this kind offer. As we plan to use the TLR4 antibody with rat and human cells we would like to use the ab91532 product as isotype control.

ANSWER:

I have issued a free of charge replacement for one vial of ab91532. To check the status of the order please contact our Customer Service team and reference this number. Please be aware this product is currently out of stock. Expect delivery around December 21. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

we are using your product nr ab7002 as isotype control for your TLR-4-antibody (nr.ab45104). As we want to identify TLR4 on monocytes we use an ED9-antibody, too. We know have a problem with the isotype control, which seems to bind to the ED-9-antibody, as you can see in the attached files. We now were wondering how this could happen, as the isotype control should not bind my ED-9-antibody. Furthermore we would like to know, what you suggest as to how we should proceed?

ANSWER:

Thank you for contacting us.

The product ab7002 is not an isotype control, but a secondary antibody that binds to antibodies raised in mouse (such as ab45104).

I am not certain which ED9-antibody (ab34086 Anti-SIRP alpha antibody [ED9] ??) you are refering to, but am not surprised that you see cross-reactivity as ab7002 is a secondary antibody and is supposed to cross react with mouse monoclonal antibodies.

A true isotype control for the TLR4 antibody (mouse monoclonal IgG2b, PE-conjugated) would be for instance:

ab91532 - Mouse IgG2b [PLPV219] (Phycoerythrin) - Isotype Control http://www.abcam.com/Mouse-IgG2b-PLPV219-Phycoerythrin-Isotype-Control-ab91532.html

ab1284 - Mouse IgG2b [HybIgG2b] (Phycoerythrin) http://www.abcam.com/Mouse-IgG2b-HybIgG2b-Phycoerythrin-ab1284.html

ab118540 - Mouse IgG2b monoclonal PE - Isotype control http://www.abcam.com/Mouse-IgG2b-monoclonal-PE-Isotype-control-ab118540.html

And a true isotype control for the ED-antibody ab34086 - if this is the antibody you are refering to - (mouse monoclonal IgG1, unconjugated) would be for instance:

ab91353 - Mouse IgG1 [ICIGG1] - Isotype Control http://www.abcam.com/Mouse-IgG1-ICIGG1-Isotype-Control-ab91353.html

ab81032 - Mouse IgG1 Monoclonal (NCG01) - Isotype Control - BSA and Azide Free http://www.abcam.com/Mouse-IgG1-Monoclonal-NCG01-Isotype-Control-BSA-and-Azide-Free-ab81032.html I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

  I would like to ask, in order to be absolutely certain, whether your secondary antibodies, mentioned below, contain Fc part: Goat F(ab')2 polyclonal Secondary Antibody to Mouse IgG - H&L (PE), pre-adsorbed (ab7002) Donkey F(ab')2 polyclonal Secondary Antibody to Mouse IgG - H&L (PE), pre-adsorbed (ab7003)   I would like to use them in the experiment where their binding to the Fc receptors would be an important issue.  

ANSWER:

Thank you for contacting us.  Our F(ab')2 fragment secondary antibodies ab7002 and ab7003 will not contain the Fc region, and this will be covered under our Abpromise guarantee.  If any Fc reactivity is observed in your experiments, we would be happy to offer you a free of charge replacement, credit, or refund.  I hope this helps, please let me know if you need any additional information. 

Thank you for the informations.

Is it possible to know approximately when you will have a new batch of this monoclonal Ab in stock?

Also, as I told you, the experiment worked well at the beginning (with the first lot# we bought) but not so well with the next lot #. My problem is, before receiving this primary antibody (second order of this same product), I had ordered an extra tube of both primary and secondary Ab because I knew I had many experiments to performed. I haven't received those Ab yet (ordered nearly 2 weeks ago) but they are probably already sent. I want to know if I can send you back those new products, knowing the Ab does not work well.

ANSWER:

Thank you for getting back to me.

If you provide me with the details of your most recent order of your primary and secondary antibodies (order number and date) I will do my best to cancel the order before it is shipped and provide you with credit to the value of the order.

If you have received the antibodies on receipt of this e-mail you can send them back in the same packaging that you received the antibodies in and you will be reimbursed (minus a 20% restocking fee). Please return them with the same ice packs that were in the box during shipping (refrozen).

I am in contact with the source of this antibody and I will be in touch when I have further information as to the availability of a new batch. I appreciate your patience in this matter.

Hi, Several weeks ago, I've bought one of your mouse monoclonal antibody (aE11 to C5b-9). As a negative control, I've also bought an isotype control (mouse IgG2a kappa monoclonal (MG2a-53)) and a Goat polyclonal antibody to Mouse IgG PE (ab7002) as a secondary antibody. The purpose of my study is to detect C5b-9 complexes on different cells (lymphocytes, monocytes, etc.) in patients with an inflammatory disease and in controls (by flowcytometry). As positive controls, I activate the complement system by different ways (so we can normally see more C5b-9 on the cells' surface when the complement is activated). At the beginning of my experiment, I've seen many significant differences between the different conditions used (patients, controls and positive controls) whereas now, I can't see those results anymore. The results I obtained with the isotype control are even higher than those with the regular monoclonal Ab; more C5b-9 complexes are present on the cells WITHOUT using the monoclonal antibody. I've noted that this event correlates with the fact that I now use new primary (lot # 166383) and secondary (154790) antibodies. I've received those new Abs less than 3 weeks ago. I first guessed that I might have diluted the secondary Ab more, so I've tried many dilutions (1:5, 1:100, 1:200): No positive results. What dilution of each Ab do you recommended to use in flowcytometry? What do you think the problem could be? Could it be because of the new Abs I use? (Which one?). What do you suggest? Is it possible to send me new Abs (different lot numbers maybe...)? Thank you for your help, Regards,

ANSWER:

Thank you for your enquiry.

I am sorry to hear that you have been having difficulties with this antibody, your enquiry definitely sounds batch related. The fact that you have been obtaining negative results in your FACS system until the arrival of the most recent batch suggests that the problem is with the new primary antibody that you have received. Unfortunately at the moment we only have batch # 166383 in stock. However, I would like to offer you a credit note that you can use against the cost of another Abcam antibody. This is provided that the antibody was purchased within the past 90 days. If this is the case please e-mail me the date of purchase and the original purchase order number.

I look forward to hearing from you.