Anti-FOXC2 抗体 (ab5060)

Thank you for your reply.

We do not batch test ab5060 in IHC on frozen sections, we only guarantee it to work in IHC-P, ChIP and western blot. Therefore that would explain why you are not getting similar results, as we do not test for this application.

Were you seeing weaker signal than before? If so have you tried to increase the primary antibody concentration? Or were you seeing high background?

Have you tried the antibody in any of the test of the guaranteed applications? If so, does the antibody work in those applications?

Thank you for contacting Abcam.

The antibody is covered under our Abpromise for six months and is guaranteed to work in WB, ChIP and IHC-P on mouse and human samples. Since you ordered it in July last year, that is outside the six month guarantee but in this case I can make an exception and say that if we cannot resolve the issue you are having with the antibody then I send a replacement antibody.

To be able to better assist and see if there are any protocol suggestions I can make, can you please send me a copy of the protocol you are using, including species/cell types, primary antibody concentrations and incubations time, any antigen retrieval performed.

I look forward to your reply and helping you to resolve this issue.

I have been able to open the reference Dagenais SL et al. Foxc2 is expressed in developing lymphatic vessels and other tissues associated with lymphedema-distichiasis syndrome. Gene Expr Patterns 4:611-9 (2004) which has used the antibody in immunohistochemistry and I hope the following detailed protocol will help you: "Five micron paraffin sections were de-paraffinized with xylene and then re-hydrated in decreasing ethanol solutions according to standard procedures. Cryosections were incubated 10 min in acetone at ?20 °C and then air dried 15 min. Sections were washed three minutes in PBS before endogenous peroxidase activity was quenched by incubation for 20 min in 1.5% hydrogen peroxide in methanol. After one three-minute wash in PBS, antigen retrieval was performed on paraffin sections. ...For Foxc2... we used high temperature retrieval by incubating 10 min in boiling 0.01 M citrate buffer (pH 6.0), which was followed by cooling for 20 min at room temperature in the acid bath. Sections were washed for 3 min in PBS and then blocked for 10 min with TNB buffer (TSA" Fluorescence Systems) containing 0.1% Triton X-100 (t-Octylphenoxypolyethoxy-ethanol; Sigma, St Louis, MO, USA). Incubation with primary antibodies was 1 h at room temperature in a humidified chamber. Incubations with secondary and tertiary antibodies were 30 min in a humidified chamber. For protein signal amplification, sections were incubated 10 min with Fluorescein Tyramide (TSA" Fluorescence Systems). All incubations were followed by 3× 3-min washes in PBS containing 0.05% Tween 20 (Sigma). After a final series washes, sections were counterstained with DAPI (4?,6-Diamidino-2-phenylindole) in Vectashield mounting medium (Vector Labs, Burlingame, CA, USA). Signals were visualized with a Zeiss Axiophot Fluorescent microscope and imaging was performed using the Olympus DP70 Digital Camera System. Standard protocols were followed for Hematoxylin & Eosin staining." Please do not hesitate to contact us if you still experience problems with this protocol,

I am delighted to hear you are satisfied with the replacement vial sent following your problem reported on the 3rd March; thank you for taking the time to let me know. Please consider submitting a review and in return we will award you 50 points with the Abcam Loyalty Scheme which can be redeemed on Amazon gift vouchers. Thank you again for your kind e-mail,

Thank you for your phone call. Originally, this product was released as a fast-track antibody and recently we have upgraded it. One of our customers has tested and used it successfully for application in IHC-formalin-fixed paraffin-embedded sections. I don't know if what you are seeing has been reported before. You may want to refer to the following publication, the authors used ab5060 in IHC: Dagenais SL et al. Foxc2 is expressed in developing lymphatic vessels and other tissues associated with lymphedema-distichiasis syndrome. Gene Expr Patterns 4:611-9 (2004). Please let me know if you require further assistance.

I'm sorry to hear you are having a problem with your recent vial of ab5060. I can certainly offer you a replacement vial if you can provide me with your order details (the Abcam order number of that particular orde and PO number) and I will arrange this as soon as possible. I look forward to hearing from you,

Thank you for your enquiry. All the information available for this product is listed on the on-line datasheet Originally, this product has been released as a fast-track antibody and recently we have upgraded it. One of our customers has tested and used it successfully for IHC application. It is good to know it works on transfectants but we can't add anything on endogenous staining because we never got any. We would highly appreciate if your customer can send us a review about this product.