MA5-33010

antibody from Invitrogen Antibodies

Targeting: FH

Western blot

Immunohistochemistry

Antibody data

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Validation data

Product number: MA5-33010 - Provider product page
Provider: Invitrogen Antibodies
Product name: Fumarase Monoclonal Antibody (9D8)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: 9D8
Vial size: 100 µg
Concentration: 500 µg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

FH protein structure - MA5-33010 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of fumarate hydratase (mitochondrial) (FH) was performed using 70% confluent log phase untreated and hydroxyurea treated (5mM for 3 hr) HCT 116 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Fumarase Monoclonal Antibody (9D8) (Product # MA5-33010) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2500), for 45 minutes at room temperature (Panel a, e: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing increased staining for FH with mitochondria-like localization in treated HCT 116 cells. Panel e represents merged image of untreated HCT 116 cells showing lower levels of staining for FH. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Fumarase (FH) was performed using 80% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Fumarase Monoclonal Antibody (9D8) (Product # MA5-33010) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing mitochondria-like localization for FH in PC-3 cells. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).