Anti-CD276 antibody [SP265] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD276 antibody [SP265] - BSA and Azide free (AB272011)

This data was developed using ab227679, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (AB272011)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227679).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD276 antibody [SP265] - BSA and Azide free (AB272011)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227679).

• WB

Lab

Western blot - Anti-CD276 antibody [SP265] - BSA and Azide free (AB272011)

This data was developed using ab227679, the same antibody clone in a different buffer formulation.

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 (<a href='/en-us/products/primary-antibodies/cd276-antibody-sp265-c-terminal-ab227679'>ab227679</a>) 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='/en-us/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='/en-us/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] - BSA and Azide free (AB272011)

This data was developed using ab227679, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-CD276 antibody [SP265] - C-terminal (<a href='/en-us/products/primary-antibodies/cd276-antibody-sp265-c-terminal-ab227679'>ab227679</a>) at 1/100 dilution

All lanes:

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

Predicted band size: 57 kDa

true