Anti-FANCI antibody [EPR23903-124]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-FANCI antibody [EPR23903-124] (AB245219)

Immunofluorescent analysis of 100% methanol-fixed HeLa cells labelling FANCI with ab245219 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear foci staining in HeLa cell line, the signal increased a little after treatment with Mitomycin C (500ng/ml) for 6h. FANCI forms nuclear foci in cells in the absence or presence of exogenous DNA damage (PMID : 19561358). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCI antibody [EPR23903-124] (AB245219)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling FANCI with ab245219 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human tonsil. The section was incubated with ab245219 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-FANCI antibody [EPR23903-124] (AB245219)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling FANCI with ab245219 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-FANCI antibody [EPR23903-124] (AB245219)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling FANCI with ab245219 at 1/600 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCI antibody [EPR23903-124] (AB245219)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling FANCI with ab245219 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human colon. The section was incubated with ab245219 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FANCI antibody [EPR23903-124] (AB245219)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling FANCI with ab245219 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining in human testis (PMID : 31219578). The section was incubated with ab245219 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 10 mins.

• WB

Lab

Western blot - Anti-FANCI antibody [EPR23903-124] (AB245219)

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

Exposure times : Lanes 1-3 : 26 seconds; Lane 4 : 125 seconds.

All lanes:

Western blot - Anti-FANCI antibody [EPR23903-124] (ab245219) at 1/5000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Jurkat (human t cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 3:

Raji ( human burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

Human testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 149 kDa

Observed band size: 149 kDa

false

• WB

Lab

Western blot - Anti-FANCI antibody [EPR23903-124] (AB245219)

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

The more slowly migrating band is a monoubiquitinated form of FANCI, which can be induced by Mitomycin C treatment.

The expression profile observed is consistent with what has been described in the literature (PMID : 17460694).

Exposure time : 70 seconds.

All lanes:

Western blot - Anti-FANCI antibody [EPR23903-124] (ab245219) at 1/2000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with 500ng/ml Mitomycin C for 6h, whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 149 kDa

Observed band size: 149 kDa

false