重组Anti-ErbB3 / HER3抗体[EPR22669-25]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling ErbB3 / HER3 with ab255607 at 1/250 dilution (2.1 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining in human urothelial carcinoma (PMID : 23333248) is observed. Counterstained with hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

Intracellular flow cytometric analysis of 4% paraformaldehydefixed 90% methanolpermeabilized MDA-MB-231 (Human breast adenocarcinoma epithelial cell, Left) / MCF7 (Human breast adenocarcinoma epithelial cell, Right) cells labeling ErbB3 / HER3 with ab255607 at 1/50 dilution (1μg)/ Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled 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. Weak expression control : MDA-MB-231 (PMID : 25018110, 16519802).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling ErbB3 / HER3 with ab255607 at 1/250 dilution (2.1 μg/ml) followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining in human breast carcinoma (PMID : 24346286) is observed. Counterstained with hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling ErbB3 / HER3 with ab255607 at 1/250 dilution (2.1 μg/ml) followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining in mouse skin (PMID : 26194695) is observed. Counterstained with hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

• WB

Lab

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

Western blot : Anti-ErbB3 / HER3 antibody [EPR22669-25] ab255607 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 90-175 kDa in Wild-type MCF7 UNBOILED cell lysates with no signal observed at this size in ERBB3 knockout MCF7 UNBOILED cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc BSA 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 were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (ab255607) at 1/1000 dilution

Lane 1:

Wild-type MCF7 UNBOILED at 20 µg

Lane 2:

Western blot - Human ERBB3 knockout MCF7 cell line (<a href='/products/cell-lines/human-erbb3-knockout-mcf7-cell-line-ab286320'>ab286320</a>) at 20 µg

Lane 3:

U-2 OS UNBOILED at 20 µg

Lane 4:

Raji UNBOILED at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 148 kDa,185 kDa

Observed band size: 90-175 kDa,76 kDa

false

• WB

Unknown

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

Fresh lysates are needed for western blot.

ErbB 3 low expression cell lines : HeLa, MDA-MB-231 (PMID : 22436610, 22301547).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (ab255607) at 1/1000 dilution

Lane 1:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) 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 1/100000 dilution

Predicted band size: 148 kDa

false

Exposure time: 26s

• WB

Unknown

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (ab255607) at 1/1000 dilution

Lane 1:

4T1 (Mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SK-BR-3 (Human breast adenocarcinoma epithelial cell) 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 1/100000 dilution

Predicted band size: 148 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (AB255607)

The expression profile/molecular weight observed is consistent with what has been described in the literature. PMID : 26194695 and 19690388.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ErbB3 / HER3 antibody [EPR22669-25] (ab255607) at 1/1000 dilution

Lane 1:

Mouse stomach lysate at 20 µg

Lane 2:

Mouse skin 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 1/20000 dilution

Predicted band size: 148 kDa

false

Exposure time: 70s