Anti-FH/Fumarase antibody [EPR21104]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

Immunohistochemical analysis of paraffin-embedded human kidney tissue (left panel) and human clear cell renal cancer tissue (right panel) labeling FH/Fumarase with ab233394 at 1/500 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 (PMID : 21695080). 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.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

Immunofluorescent analysis of 100% methanol-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling FH/Fumarase with ab233394 at 1/100 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.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling FH/Fumarase with ab233394 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled 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

• IP

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Immunoprecipitation - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

FH/Fumarase was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab233394 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233394 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input).

Lane 2 : ab233394 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab233394 in HeLa whole cell lysate.

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-FH/Fumarase antibody [EPR21104] (ab233394)

Predicted band size: 54 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling FH/Fumarase with ab233394 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular staining on 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.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling FH/Fumarase with ab233394 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Granular staining on rat 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.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

Immunofluorescent analysis of 100% methanol-fixed NIH/3T3 (mouse embyro fibroblast cell line) cells labeling FH/Fumarase with ab233394 at 1/100 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.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

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

• WB

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Western blot - Anti-FH/Fumarase antibody [EPR21104] (AB233394)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lanes 1-5 : 26 seconds; Lanes 5-6 : 10 seconds; Lane 8-10 : 3 minutes.

All lanes:

Western blot - Anti-FH/Fumarase antibody [EPR21104] (ab233394) at 1/1000 dilution

Lane 1:

HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 3:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate at 10 µg

Lane 5:

C6 (rat glial tumor cell line) whole cell lysate at 10 µg

Lane 6:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 7:

HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 10 µg

Lane 8:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 9:

Human brain lysate at 10 µg

Lane 10:

Human fetal kidney lysate at 10 µg

Secondary

Lanes 1 - 8:

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

Lanes 9 - 10:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/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

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