Anti-SON/DBP5 antibody [EPR28617-69]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SON/DBP5 antibody [EPR28617-69] (AB317311)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A431(human epidermoid carcinoma epithelial cell) cells labelling SON/DBP5 with ab317311 at 1/500 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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SON/DBP5 antibody [EPR28617-69] (AB317311)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling SON/DBP5 with ab317311 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon carcinoma. The section was incubated with ab317311 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SON/DBP5 antibody [EPR28617-69] (AB317311)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling SON/DBP5 with ab317311 at 1/2000 (0.254 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear staining in A431 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). 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 (2 ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SON/DBP5 antibody [EPR28617-69] (AB317311)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling SON/DBP5 with ab317311 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human testis. The section was incubated with ab317311 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-SON/DBP5 antibody [EPR28617-69] (AB317311)

SON can undergo degradation easily.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18952841, 27545680).

Lysates of lanes 2-4 were freshly made and used for Western Blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-SON/DBP5 antibody [EPR28617-69] (ab317311) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A431 whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 264 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-SON/DBP5 antibody [EPR28617-69] (AB317311)

SON can undergo degradation easily.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18952841, 27545680).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SON/DBP5 antibody [EPR28617-69] (ab317311) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

HeLa transfected with siRNA specifically targeting SON whole cell lysate at 40 µg

Secondary

All lanes:

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

Observed band size: 264 kDa,124 kDa

false

Exposure time: 26s