Anti-MEF2D 抗体 [EPR24993-10]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized THP-1 cells labelling MEF2D with ab282731 at 1/50 (11.42 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing mainly nuclear staining in THP-1 cells is observed.

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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) cells labelling MEF2D with ab282731 at 1/500 dilution (0.1 ug) (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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Nuclear staining on rat colon. The section was incubated with ab282731 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) .

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Nuclear staining on rat cererbum (PMID : 28738418). The section was incubated with ab282731for 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) .

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Nuclear staining on mouse testis (PMID : 24694307). The section was incubated with ab282731 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) .

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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling MEF2D with ab282731 at 1/500 dilution (0.1 ug) (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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Nuclear staining on mouse cererbum (PMID : 28738418). The section was incubated with ab282731 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) .

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling MEF2D with ab282731 at 1/50 (11.42 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green) Confocal image showing mainly nuclear staining in RAW 264.7 cells is observed.

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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

Lab

Western blot - Anti-MEF2D antibody [EPR24993-10] (AB282731)

False colour image of Western blot : Anti-MEF2D antibody [EPR24993-10] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab282731 was shown to bind specifically to MEF2D. A band was observed at 67 kDa in wild-type HeLa cell lysates with no signal observed at this size in mef2d CRISPR-Cas9 edited cell line ab281636. The band observed in the CRISPR-Cas9 edited lysate lane below 67 kDa is likely to represent a truncated form of MEF2D. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and mef2d CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used

Lanes 1 - 4:

Western blot - Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-MEF2D antibody [EPR24993-10] - BSA and Azide free (<a href='/products/primary-antibodies/mef2d-antibody-epr24993-10-bsa-and-azide-free-ab282733'>ab282733</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MEF2D knockout HeLa cell line (ab281636)

Lane 2:

mef2d CRISPR-Cas9 edited HeLa cell lysate at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

NIH/3T3 cell lysate at 20 µg

Secondary

All lanes:

oat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 67 kDa

false

• WB

Lab

Western blot - Anti-MEF2D antibody [EPR24993-10] (AB282731)

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

His-tagged human MEF2D recombinant protein (aa54-328) 10ng

Myc/DDDDK-tagged human MEF2A recombinant protein 10ng

Myc/DDDDK-tagged human MEF2B recombinant protein 10ng

Myc/DDDDK-tagged human MEF2C recombinant protein 10ng

Exposure time : 10 seconds

All lanes:

Western blot - Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution

Lane 1:

His-tagged human MEF2D recombinant protein at 0.01 µg

Lane 2:

Myc/DDDDK-tagged human MEF2A recombinant protein at 0.01 µg

Lane 3:

Myc/DDDDK-tagged human MEF2B recombinant protein at 0.01 µg

Lane 4:

Myc/DDDDK-tagged human MEF2C recombinant protein at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 56 kDa

Observed band size: 31 kDa

false

• WB

Lab

Western blot - Anti-MEF2D antibody [EPR24993-10] (AB282731)

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

Low expression : SW480/ DLD-1 (PMID : 27364559).

Exposure time : 147 seconds

All lanes:

Western blot - Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution

Lane 1:

THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 20 µg

Lane 2:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

DLD-1 (human colon epithelial) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 56 kDa

Observed band size: 40 kDa,70 kDa

false

• WB

Lab

Western blot - Anti-MEF2D antibody [EPR24993-10] (AB282731)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTLysates were made freshly and used in WB test immediately to minimize protein degradation.

Exposure time : 147 seconds

All lanes:

Western blot - Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

2.4G2 (rat B cell lymphoma B lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 56 kDa

Observed band size: 70 kDa

false