Anti-FMO3 antibody [EPR6968]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FMO3 antibody [EPR6968] (AB126711)

Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling FMO3 with purified ab126711 at 1/150. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500) were also used.

Control 1 : primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMO3 antibody [EPR6968] (AB126711)

Immunohistochemitsy (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling FMO3 with unpurified ab126711 at a 1/500 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMO3 antibody [EPR6968] (AB126711)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling FMO3 with purified ab126711 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMO3 antibody [EPR6968] (AB126711)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling FMO3 with purified ab126711 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FMO3 antibody [EPR6968] (AB126711)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling FMO3 with purified ab126711 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

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Western blot - Anti-FMO3 antibody [EPR6968] (AB126711)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FMO3 antibody [EPR6968] (ab126711) at 1/5000 dilution

Lane 1:

Human fetal kidney tissue lysate at 20 µg

Lane 2:

Human fetal liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 60 kDa

Observed band size: 56 kDa

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• WB

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Western blot - Anti-FMO3 antibody [EPR6968] (AB126711)

All lanes:

Western blot - Anti-FMO3 antibody [EPR6968] (ab126711) at 1/1000 dilution

Lane 1:

Human fetal liver tissue lysate at 10 µg

Lane 2:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 60 kDa

false

• WB

Unknown

Western blot - Anti-FMO3 antibody [EPR6968] (AB126711)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FMO3 antibody [EPR6968] (ab126711) at 1/5000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Rat kidney tissue lysate at 20 µg

Lane 4:

Rat liver tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 60 kDa

Observed band size: 56 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-FMO3 antibody [EPR6968] (AB126711)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• WB

CiteAb

Western blot - Anti-FMO3 antibody [EPR6968] (AB126711)

FMO3 western blot using anti-FMO3 antibody [EPR6968] ab126711. Publication image and figure legend from Li, L., Chen, B., et al., 2019, Aging (Albany NY), PubMed 31715585.

ab126711 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab126711 please see the product overview.

Fructus Ligustri Lucidi (FLL) decreases the expression of FMO3 and the levels of circulating TMAO in aging mice. (A) The circulating TMAO levels quantified by LC/MS/MS. The expression levels of FMO3 were analyzed by immunohistochemical staining (B–C) and western blot (D) in the livers. (E–F) Correlation heat map indicating the association between the specific microbiota taxonomic genera and FMO3 expressions (E) and TMAO levels (F). Red indicates a positive association, blue denotes a negative association, and white manifests no association. a denotes a significant false discovery rate (FDR)-adjusted positive correlation at p values of < 0.05, and aa denotes a significant FDR-adjusted positive correlation at p values of < 0.01. b denotes a significant FDR-adjusted negative correlation at p values of < 0.05, and bb denotes a significant FDR-adjusted negative correlation at P values of < 0.01. Data are presented as mean ± SD. # compared with the normal group. * compared with the aging model group. p < 0.05 was considered statistical difference, n=8.

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