Overview

  • Product name

    Anti-FH/Fumarase antibody [8F12BB5]
    See all FH/Fumarase primary antibodies
  • Description

    Mouse monoclonal [8F12BB5] to FH/Fumarase
  • Host species

    Mouse
  • Tested applications

    Suitable for: In-Cell ELISA, IP, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Rat, Cow, Human
  • Immunogen

    Full length native protein (purified). This information is considered to be commercially sensitive.

  • Positive control

    • Human HDFn cells; Human cerebellum tissue; HepG2 cells, human liver mitochondria, bovine liver mitochondria, and rat liver mitochondria; HeLa cells
  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

     This product was previously labelled as FH

     

Properties

Applications

Our Abpromise guarantee covers the use of ab110286 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
In-Cell ELISA Use a concentration of 4 µg/ml. 0.4 µg/well
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval - 1 min pressure cook in 1mmol EDTA pH8.
ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use a concentration of 1 µg/ml.

Recommend 0.1% Triton X-100 permeabilization.

 

 

 

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Also acts as a tumor suppressor.
  • Pathway

    Carbohydrate metabolism; tricarboxylic acid cycle; (S)-malate from fumarate: step 1/1.
  • Involvement in disease

    Defects in FH are the cause of fumarase deficiency (FHD) [MIM:606812]; also known as fumaricaciduria. FHD is characterized by progressive encephalopathy, developmental delay, hypotonia, cerebral atrophy and lactic and pyruvic acidemia.
    Defects in FH are the cause of multiple cutaneous and uterine leiomyomata (MCUL1) [MIM:150800]. MCUL1 is an autosomal dominant condition in which affected individuals develop benign smooth muscle tumors (leiomyomata) of the skin. Affected females also usually develop leiomyomata of the uterus (fibroids).
    Defects in FH are the cause of hereditary leiomyomatosis and renal cell cancer (HLRCC) [MIM:605839].
  • Sequence similarities

    Belongs to the class-II fumarase/aspartase family. Fumarase subfamily.
  • Cellular localization

    Cytoplasm and Mitochondrion.
  • Information by UniProt
  • Database links

  • Alternative names

    • FH antibody
    • Fumarase antibody
    • Fumarate hydratase antibody
    • Fumarate hydratase mitochondrial antibody
    • Fumarate hydratase, mitochondrial antibody
    • FUMH_HUMAN antibody
    • HLRCC antibody
    • LRCC antibody
    • MCL antibody
    • MCUL 1 antibody
    • MCUL1 antibody
    • MS709 antibody
    • Multiple hereditary cutaneous leiomyomata antibody
    see all

Images

  • HeLa cells were stained with 1 µg/mL ab110286 (blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
  • Immunocytochemistry image of FH/Fumarase (ab110286) stained human HDFn cells. The cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15min). The cells were incubated with ab110286 at 5µg/ml for 2h at room temperature or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps. DAPI was used to stain the cell nuclei (blue). Target protein locates mainly in mitochondria.

  • FH/Fumarase immunocaptured from HepG2 cells (Lane 1), human liver mitochondria (Lane 2), bovine liver mitochondria (Lane 3), and rat liver mitochondria (Lane 4) using ab110286.

  • ab110286 staining FH/Fumarase in formalin-fixed, paraffin-embedded human cerebellum tissue by Immunohistochemistry at 5ug/ml. FH/Fumarase immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells.

References

This product has been referenced in:

  • Chen T  et al. PAK4 phosphorylates fumarase and blocks TGF-ß-induced cell growth arrest in lung cancer cells. Cancer Res N/A:N/A (2019). Read more (PubMed: 30683654) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question

Thank you very much for your attention to the problems we are facing with the antibody ab 110286 in our immunohistochemical staining protocol. Meanwhile, I have carried out more experiments which also gave unsuccessful results. I am summarizing the information you requested and attaching a table with all the variables we have been introducing to the protocol mainly concerning the antigen retrieval and primary antibody conditions.

I hope you find this information helpful and I am looking forward to hearing from as soon as possible.

Many wishes,


1. Sample type, species, tissue: Human kidney and human liver, paraffin embedded tissue sections (2-3µ);

2. Sample fixation: 10% neutral buffered formalin

3. Antigen retrieval, permeabilization: samples were manually deparaffinised (clear rite and ethanol 100%, 95% and 70%). We tried different antigen retrieval approaches, please report to the attached table;

4. Blocking conditions: Lab Vision Corporation Thermo Scientific: (1) Avidin/Biotin blocking system, (2) Streptavidin peroxidase and (3) Ultra V block;

5. Primary antibody conditions: please see attached table;

6. Secondary antibody conditions: biotinylated goat polyvalent antibody (anti mouse and anti-rabbit), Lab vision Corporation Thermo Scientific;

7. Wash steps: we are using in house prepared phosphate- tween 20 buffer , pH 7.4 for all IHC-P protocols

8. Detection method, amplification steps: DAB chromogen and substrate buffer, Dako;

9. Controls used: we used human normal liver and human kidney as positive control, matched samples were used as negative controls by replacing the primary antibody with Ultra antibody diluent, Lab Vision Corporation Thermo Scientific in all experiments.

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Answer

Thank you for your response and for passing some useful experimental details. Your co-operation is very much appreciated in this matter.

I can see the several attempts have been made to optimize the staining. The protocol looks fine to me and at this stage I can conclude that the antibody may have been accidentally damaged during shipment or storage.

I can offer you a new vial as a free of charge replacement or a credit note which you can use in the future for any of our products which are in the catalogue. Please do let me know how you wish to proceed with your enquiry.

I look forward to hearing from you soon.

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Answer

Thank you for contacting us.

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



Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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