Anti-MDC1 antibody [EPR24360-116]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDC1 antibody [EPR24360-116] (AB271061)

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labelling MDC1 with ab271061 at 1/500 (1.046 μg/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Nuclear staining on human ovarian cancer. The section was incubated with ab271061 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 LeicaDS9800 (Bond® Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MDC1 antibody [EPR24360-116] (AB271061)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293 (Human embryonic kidney epithelial cell) cells labelling MDC1 with ab271061 at 1/500 dilution (0.1μg)/ red (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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDC1 antibody [EPR24360-116] (AB271061)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling MDC1 with ab271061 at 1/500 (1.046 μg/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Nuclear staining on human testis. The section was incubated with ab271061 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 LeicaDS9800 (Bond® Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MDC1 antibody [EPR24360-116] (AB271061)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling MDC1 with ab271061 at 1/50 (10.46 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2μg/ml) dilution (Green). Confocal image showing nuclear staining in HeLa cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2μg/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MDC1 antibody [EPR24360-116] (AB271061)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling MDC1 with ab271061 at 1/500 dilution (0.1μg)/ red (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, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-MDC1 antibody [EPR24360-116] (AB271061)

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

Fresh lysates were used in this WB.

This blot was developed using a higher sensitivity ECL substrate.

The expression profile/ molecular weight observed is consistent with what has been described in the literature ( PMID : 12607005; PMID : 26701181 )

Exposure time : 3 minutes

All lanes:

Western blot - Anti-MDC1 antibody [EPR24360-116] (ab271061) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Hela transfected with MDC1 siRNA 1 whole cell lysate at 20 µg

Lane 3:

Hela transfected with MDC1 siRNA 2 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: 227 kDa

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