FH Rabbit Polyclonal Antibody
cat.: ER1901-10
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 54 kDa
Isotype: IgG
Immunogen: Recombinant protein within human Fumarase aa 83-267.
Positive control: SH-SY5Y cell lysates, mouse colon tissue lysates, rat colon tissue lysates, SW620, Siha, A431, rat testis tissue, human liver tissue, human liver carcinoma tissue, human kidney tissue, mouse small intestine tissue, HCT116, LO2 cell lysate, mouse liver tissue lysate, rat liver tissue lysate.
Subcellular location: Mitochondrion, cytosol, nucleus, chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:2,000
1:50-1:100
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P07954 Human | P97807 Mouse | P14408 Rat
Alternative names: FH Fumarase Fumarate hydratase Fumarate hydratase mitochondrial Fumarate hydratase, mitochondrial FUMH_HUMAN HLRCC LRCC MCL MCUL 1 MCUL1 MS709 Multiple hereditary cutaneous leiomyomata
Images
ER1901-10_1.jpg Fig1: Western blot analysis of FH on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1901-10, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: SH-SY5Y cell lysate
Lane 2: Mouse colon tissue lysate
Lane 3: Rat colon tissue lysate
ER1901-10_2.png Fig2: ICC staining of FH in SW620 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1901-10, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1901-10_3.png Fig3: Immunocytochemistry analysis of Siha cells labeling FH with Rabbit anti-FH antibody (ER1901-10) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-FH antibody (ER1901-10) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
ER1901-10_4.png Fig4: Immunocytochemistry analysis of A431 cells labeling FH with Rabbit anti-FH antibody (ER1901-10) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-FH antibody (ER1901-10) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
ER1901-10_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-FH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-10, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1901-10_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-FH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-10, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1901-10_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-FH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-10, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1901-10_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-FH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-10, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1901-10_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue using anti-FH antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1901-10, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1901-10_10.jpg Fig10: Flow cytometric analysis of FH was done on HCT116 cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1901-10, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ER1901-10_11.jpg Fig11: Western blot analysis of FH on different lysates with Rabbit anti-FH antibody (ER1901-10) at 1/1,000 dilution.

Lane 1: LO2 cell lysate (10 µg/Lane)
Lane 2: Mouse liver tissue lysate (20 µg/Lane)
Lane 3: Rat liver tissue lysate (20 µg/Lane)

Predicted band size: 55 kDa
Observed band size: 50 kDa

Exposure time: 1 minute;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER1901-10) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.