Recombinant
RabMAb

Recombinant Anti-FH/Fumarase antibody [EPR21105] (ab233393)

Overview

  • Product name

    Anti-FH/Fumarase antibody [EPR21105]
    See all FH/Fumarase primary antibodies
  • Description

    Rabbit monoclonal [EPR21105] to FH/Fumarase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human FH/Fumarase aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: P07954

  • Positive control

    • WB: HeLa, HEK-293, Jurkat, NIH/3T3 and PC-12 whole cell lysates; Rat brain and spleen tissue lysates; human fetal brain and fetal spleen tissue lysates. IHC-P: Human kidney and clear cell renal cancer tissue; mouse kidney tissue; rat kidney tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow cyt: HeLa cells. IP: HEK-293 whole cell lysate.
  • General notes

     This product was previously labelled as FH

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab233393 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 1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 54 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
Flow Cyt 1/500.
IP 1/30.

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

  • Lanes 1-7 : Anti-FH/Fumarase antibody [EPR21105] (ab233393) at 1/5000 dilution
    Lanes 8-9 : Anti-FH/Fumarase antibody [EPR21105] (ab233393) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
    Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
    Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
    Lane 4 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg
    Lane 5 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg
    Lane 6 : Rat brain tissue lysate at 10 µg
    Lane 7 : Rat spleen tissue lysate at 10 µg
    Lane 8 : Human fetal brain tissue lysate at 10 µg
    Lane 9 : Human fetal spleen tissue lysate at 10 µg

    Secondary
    Lanes 1-7 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 8-9 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Developed using the ECL technique.

    Predicted band size: 54 kDa
    Observed band size: 48 kDa
    why is the actual band size different from the predicted?



    Exposure times:

    Lanes 1-5: 6 seconds.
    Lanes 6 & 7: 8 seconds.
    Lanes 8 & 9: 3 seconds.

    Blocking and dilution buffer: 5% NFDM/TBST.

    The lower mass bands could represent isoforms which have been described in the literature (PMID: 434773)

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling FH/Fumarase with ab233393 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling FH/Fumarase with ab233393 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Granular, cytoplasmic and nuclear staining on rat kidney (PMID: 17111171) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling FH/Fumarase with ab233393 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Intensive granular and weak nuclear staining in mouse kidney is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • FH/Fumarase was immunoprecipitated from 0.35 mg of HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate with ab233393 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233393 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Lane 1: HEK-293 whole cell lysate 10 μg (Input).
    Lane 2: ab233393 IP in HEK-293 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab233393 in HEK-293 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 10 seconds.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling FH/Fumarase with ab233393 at 1/500 dilution (red) compared with an Isotype control details (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling FH/Fumarase with ab233393 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue (left) and human clear cell renal cancer tissue (right) tissue labeling FH/Fumarase with ab233393 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Intense granular staining on human kidney whereas weaker staining in clear cell renal cancer tissue (PMID: 21695080) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

References

ab233393 has not yet been referenced specifically in any publications.

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