Overview

  • Product name
    Anti-FOXC2 antibody - ChIP Grade
    See all FOXC2 primary antibodies
  • Description
    Goat polyclonal to FOXC2 - ChIP Grade
  • Host species
    Goat
  • Specificity
    The antibody labels FOXC2 in mouse lymphatic primordia, jugular lymph sacs, lymphatic collectors and capillaries. In western blotting the antibody has been tested on FOXC2 overexpressing cell lines but not on endogenous protein.
  • Tested applications
    Suitable for: WB, ChIP, IHC-Pmore details
    Unsuitable for: IHC-Fr
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Dog
  • Immunogen

    Synthetic peptide:

    RHAAPYSYDCTKY

    , corresponding to C terminal amino acids 489-501 of Human FOXC2.

  • Positive control
    • IHC: mouse lymphatic primordia, jugular lymph sacs, lymphatic collectors and capillaries WB: FOXC2 overexpressing cell lines

Properties

Applications

Our Abpromise guarantee covers the use of ab5060 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 0.1 - 0.2 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 56 kDa).Can be blocked with Human FOXC2 peptide (ab23061).
ChIP Use at an assay dependent concentration. PubMed: 18579532
IHC-P Use a concentration of 2 - 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for IHC-Fr.
  • Target

    • Function
      Transcriptional activator. Might be involved in the formation of special mesenchymal tissues.
    • Involvement in disease
      Defects in FOXC2 are the cause of lymphedema hereditary type 2 (LMPH2) [MIM:153200]; also known as Meige lymphedema. Hereditary lymphedema is a chronic disabling condition which results in swelling of the extremities due to altered lymphatic flow. Patients with lymphedema suffer from recurrent local infections, and physical impairment.
      Defects in FOXC2 are a cause of lymphedema-yellow nails (LYYN) [MIM:153300]. LYYN is characterized by yellow, dystrophic, thick and slowly growing nails, associated with lymphedema and respiratory involvement. Lymphedema occurs more often in the lower limbs. It can appear at birth or later in life. Onset generally follows the onset of ungual abnormalities.
      Defects in FOXC2 are a cause of lymphedema-distichiasis (LYD) [MIM:153400]. LYD is characterized by primary limb lymphedema usually starting at puberty (but in some cases later or at birth) and associated with distichiasis (double rows of eyelashes, with extra eyelashes growing from the Meibomian gland orifices).
    • Sequence similarities
      Contains 1 fork-head DNA-binding domain.
    • Cellular localization
      Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • Drosphilia Forkhead Homolog Like 14 antibody
      • FKHL 14 antibody
      • FKHL14 antibody
      • Forkhead Box C2 antibody
      • Forkhead box protein C2 antibody
      • Forkhead related protein FKHL14 antibody
      • Forkhead-related protein FKHL14 antibody
      • FOX C2 antibody
      • Foxc2 antibody
      • FOXC2_HUMAN antibody
      • LD antibody
      • Mesenchyme fork head protein 1 antibody
      • Mesenchyme Forkhead 1 antibody
      • MFH 1 antibody
      • MFH 1 protein antibody
      • MFH-1 protein antibody
      • MFH1 antibody
      • Transcription factor FKH 14 antibody
      • Transcription factor FKH-14 antibody
      see all

    Images

    • ab5060 (2µg/ml) staining of paraffin embedded Human Breast cancer tissue shows nuclear staning with some weak cytoplasm staining. Steamed antigen retrieval with citrate buffer pH 6, HRP-staining.

    • FoxC2 is highly expressed in COS7 and 293 cells transiently transfected with pcDNA-FOXC2. This figure shows a western blot incubated with the ab5060 antibody after running lysates from COS7 and 293 cells transiently expressing FoxC2 (lane 1 and 3 respectively). Lane 2 shows a negative control of 293 cells transfected with an empty vector. Review by Michal Bell submitted 19 August 2004

      FoxC2 is highly expressed in COS7 and 293 cells transiently transfected with pcDNA-FOXC2. This figure shows a western blot incubated with the ab5060 antibody after running lysates from COS7 and 293 cells transiently expressing FoxC2 (lane 1 and 3 respectively). Lane 2 shows a negative control of 293 cells transfected with an empty vector. Review by Michal Bell submitted 19 August 2004
    • Anti-FOXC2 antibody - ChIP Grade (ab5060) at 1 µg/ml + Human breast cancer lysates at 30 µg

      Developed using the ECL technique.

      Predicted band size: 56 kDa
      Observed band size: 52 kDa
      why is the actual band size different from the predicted?



      The primary antibody incubated for 1 hour.

    References

    This product has been referenced in:
    • Trost A  et al. Lymphatic markers in the human optic nerve. Exp Eye Res 173:113-120 (2018). IHC ; Human . Read more (PubMed: 29746818) »
    • Fatima A  et al. Foxc1 and Foxc2 deletion causes abnormal lymphangiogenesis and correlates with ERK hyperactivation. J Clin Invest 126:2437-51 (2016). ChIP ; Mouse . Read more (PubMed: 27214551) »
    See all 19 Publications for this product

    Customer reviews and Q&As

    1-7 of 7 Abreviews or Q&A

    Answer

    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?

    Read More

    Answer

    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.

    Read More

    Answer

    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,

    Read More

    Answer

    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,

    Read More

    Answer

    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.

    Read More

    Answer

    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,

    Read More

    Answer

    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.

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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