重组Anti-B7H4抗体[EPR20236]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (AB209242)

Immunohistochemical analysis of paraffin-embedded sections in Human endometrial adenocarcinoma tissue labelling B7H4 with ab209242 at 1/1000 dilution, followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) and LeicaDS9800 (Bond™ Polymer Refine Detection). Counter stained with Hematoxylin.

A : Paraffin-embedded tissue section was incubated with ab209242 at +4 ℃ overnight. B : Paraffin-embedded tissue section was incubated with ab209242 for 30 minutes at room temperature. Immunostaining was performed on a Leica Biosystems BOND® RX instrument.

A : Heat mediated antigen retrieval using ab93678(Citrate buffer, pH 6.0) B : Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution1) for 20 mins

The two images were generated at different times and are provided to illustrate the variation in staining patterns observed between different tumor samples. For example, in endometrial adenocarcinoma, samples from different patients or tumor stages may show markedly different staining results.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-B7H4 antibody [EPR20236] (AB209242)

Immunofluorescent analysis of 100% methanol-fixed SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling B7H4 with ab209242 at 1/250 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on SK-BR-3 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (AB209242)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling B7H4 with ab209242 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Predominantly apical luminal membranous staining on human breast epithelium is observed [PMID : 15756008]. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (AB209242)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling B7H4 with ab209242 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous staining on distal convoluted renal tubules of human kidney is observed [PMID : 16798883]. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B7H4 antibody [EPR20236] (AB209242)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling B7H4 with ab209242 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous and weak cytoplasmic staining on tumor cells of human breast cancer is observed [PMID : 15756008]. Counter stained with Hematoxylin.

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

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-B7H4 antibody [EPR20236] (AB209242)

Blocking/Dilution buffer : 5% NFDM/TBST.

B7H4 is N-glycosylated protein. The level of B7H4 glycosylation varies between different cells and tissues. The bands at 65kDa and 45kDa represent glycosylated B7H4. The bands at 28kDa and 18kDa represent de-glycosylated B7H4. HT-29 and HeLa cell lines are negative controls. (PMID : 15878339).

All lanes:

Western blot - Anti-B7H4 antibody [EPR20236] (ab209242) at 1/1000 dilution

Lane 1:

SK-BR-3 (Human mammary gland adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Human breast cancer lysate at 20 µg

Lane 4:

HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 5:

HeLa (Human epithelial cell line from cervix adenocarcinoma) 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: 31 kDa

Observed band size: 18 kDa,28 kDa,45 kDa,65 kDa

false

Exposure time: 30s