Donkey polyclonal Secondary Antibody to Goat IgG - H&L (FITC) (ab6881)

Thank you for your responce I have attached the protocol that we use

The lot number for the secondary was lot63139 and the cat no. it ab6881-1 I do not know the exact time it took for the order but the product was order by the Brombacher lab at University of Cape Town, Infectious disease unit

Please lte me know if you need more info.

Thanks for your time

ANSWER:

Thank you for your protocol and your order information, we have sold only 1 vial of anti-Foxp3 to a customer in Cape town but the address is very different and it was bought in March, could this be you?

I have carefully looked at each step of your protocol and would recommend the following modifications:

-you say "defrost after fixing" and then mention fixing again. Can I please make sure the sections are fresh frozen and not perfusion fixed. Fresh frozen sections should be then postfixed for 20min in 4% paraformaldehyde in PB, or for 10min in ice cold acetone.

-I recommend washing slides in PBS 3-4 times between each step, to decrease non specific binding

-use the dilution buffer PBST to dilute the blocking agent and antibodies. PBST is made of PBS + 0.3% v/v TritonX100, to permeabilise the cells. Even if your sections are very thin they may need permeabilisation (if you fix with acetone this is not necessary as acetone fixes and permeabilises too)

-block in normal serum 10% rather than BSA, this can give problems in some rare cases

- try to use biotin-ABC-DAB rather than IF, as this is more specific and more sensitive.

I hope these suggestions will help you, please let me know how you get on and good luck with your experiments,

Need information on:

FOXP3 antibody cat. ab2481 Abcam

Goat IgG antibody (ab6881) datasheet 1 mg cat. ab6881 Abcam

I need to know what kind of resultS can i obtain if we use them with PERCP or SYCRON .

I dont see any information in your website ... We hope you can help me with this please If you have any questions please feel to call at any time,,

ANSWER:

Thank you for your email. For PERCP, do you mean the tandem conjugate PerCP-Cy5.5? I'm not sure what you mean by Sycron so it would great if you could provide me with some more information.