Recombinant Anti-FH/Fumarase antibody [EPR21105] (ab233393)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21105] to FH/Fumarase
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Reacts with: Mouse, Rat, Human
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
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21105 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
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
- Entrez Gene: 2271 Human
- Entrez Gene: 14194 Mouse
- Entrez Gene: 24368 Rat
- Omim: 136850 Human
- SwissProt: P07954 Human
- SwissProt: P97807 Mouse
- SwissProt: P14408 Rat
- Unigene: 592490 Human
see all -
Alternative names
- FH antibody
- Fumarase antibody
- Fumarate hydratase 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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FH/Fumarase antibody [EPR21105] (ab233393)
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).
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FH/Fumarase antibody [EPR21105] (ab233393)
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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FH/Fumarase antibody [EPR21105] (ab233393)
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).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
Certificate of Compliance
References (0)
ab233393 has not yet been referenced specifically in any publications.