Anti-CD276 抗体 [SP265] - C-terminal

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD276 antibody [SP265] - C-terminal (AB227679)

Flow cytometric overlay histogram analysis of 4% paraformaldehyde cells labelling CD276 KO HEK293T (ED040783) (Magenta and grey) cells permeabilised with 90% methanol and Parental HEK293T (EDWT04) (Green and black) cells permeabilised with 90% methanol labelling CD276 with ab227679 at 1/20 dilution (Magenta and Green) compared with a Rabbit monoclonal IgG (ab172730) (Black and Grey) Isotype control. Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150081), at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CD276 antibody [SP265] - C-terminal (AB227679)

Immunocytochemistry/ Immunofluorescence analysis of LNCaP (Human prostate carcinoma epithelial cell) cells labeling CD276 with purified ab227679 at 1/25 (9.24 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488 ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD276 antibody [SP265] - C-terminal (AB227679)

Flow cytometric analysis of PC-3 (human prostate adenocarcinoma cell line) cell line labeling CC276 with ab227679 at 1/400 (green) compared with a rabbit isotype control (blue).

• WB

Lab

Western blot - Anti-CD276 antibody [SP265] - C-terminal (AB227679)

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

Performed under reducing conditions.

In Western blot, ab227679 was shown to bind specifically to CD276. Target of interest was observed at 90-100 kDa in wild-type HEK-293T cell lysates (lane 1) with no signal observed at this size in CD276 knockout cell line (lane 2) (lane 2, knockout cell line ab266658.

All lanes:

Western blot - Anti-CD276 antibody [SP265] - C-terminal (ab227679) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 50 µg

Lane 2:

CD276 knockout HEK-293T whole cell lysate at 50 µg

Lane 2:

Western blot - Human CD276 knockout HEK-293T cell line (<a href='/products/cell-lines/human-cd276-knockout-hek-293t-cell-line-ab266658'>ab266658</a>) at 50 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

Observed band size: 90-110 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-CD276 antibody [SP265] - C-terminal (AB227679)

All lanes:

Western blot - Anti-CD276 antibody [SP265] - C-terminal (ab227679) at 1/100 dilution

All lanes:

Hs 700T (human pancreatic adenocarcinoma cell line) cell lysate

Predicted band size: 57 kDa

true