Anti-FH/Fumarase antibody [EPR21105]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-FH/Fumarase antibody [EPR21105] (AB233393)

Intracellular flow cytometric analysis of4% 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FH/Fumarase antibody [EPR21105] (AB233393)

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.

• IHC-P

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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).

• IP

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Immunoprecipitation - Anti-FH/Fumarase antibody [EPR21105] (AB233393)

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.

All lanes:

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

Predicted band size: 54 kDa

Observed band size: 45 kDa

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• IHC-P

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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).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FH/Fumarase antibody [EPR21105] (AB233393)

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.

• IHC-P

Supplier Data

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).

• WB

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Western blot - Anti-FH/Fumarase antibody [EPR21105] (AB233393)

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)

Lanes 1 - 7:

Western blot - Anti-FH/Fumarase antibody [EPR21105] (ab233393) at 1/5000 dilution

Lanes 8 - 9:

Western blot - 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:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lanes 8 - 9:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/4000 dilution

Predicted band size: 54 kDa

Observed band size: 48 kDa

true