Anti-SP3 antibody - N-terminal

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP3 antibody - N-terminal (AB227856)

Paraffin-embedded human ovarian carcinoma tissue stained for SP3 using ab227856 at 1/1000 dilution in immunohistochemical analysis.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP3 antibody - N-terminal (AB227856)

Paraffin-embedded human breast carcinoma tissue stained for SP3 using ab227856 at 1/1000 dilution in immunohistochemical analysis.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SP3 antibody - N-terminal (AB227856)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for SP3 (green) using ab227856 at 1/500 dilution in ICC/IF. Cells were fixed in 4% paraformaldehyde at RT for 5 minutes.

Blue : Hoechst 33342 staining.

• IP

Supplier Data

Immunoprecipitation - Anti-SP3 antibody - N-terminal (AB227856)

SP3 was immunoprecipitated from Jurkat (human T cell leukemia cell line from peripheral blood) whole cell extract with 5 μg ab227856. Western blot was performed from the immunoprecipitate using ab227856. Anti-Rabbit IgG was used as a secondary reagent.

Lane 1 : Jurkat whole cell extract.

Lane 2 : Control IgG IP in Jurkat whole cell extract.

Lane 3 : ab227856 IP in Jurkat whole cell extract.

All lanes:

Immunoprecipitation - Anti-SP3 antibody - N-terminal (ab227856)

Predicted band size: 82 kDa

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• WB

Lab

Western blot - Anti-SP3 antibody - N-terminal (AB227856)

False colour image of Western blot : Anti-SP3 antibody - N-terminal staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab227856 was shown to bind specifically to SP3. A band was observed at 100 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in SP3 knockout cell line ab266879 (knockout cell lysate ab257700). To generate this image, wild-type and SP3 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk 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 IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-SP3 antibody - N-terminal (ab227856) at 1/500 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

SP3 knockout HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human SP3 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-sp3-knockout-hct116-cell-line-ab266879'>ab266879</a>)

Predicted band size: 82 kDa

Observed band size: 100 kDa

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• WB

Supplier Data

Western blot - Anti-SP3 antibody - N-terminal (AB227856)

7.5 % SDS-PAGE gel.

All lanes:

Western blot - Anti-SP3 antibody - N-terminal (ab227856) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell extract at 30 µg

Lane 2:

Raji (human Burkitt's lymphoma cell line) whole cell extract at 30 µg

Lane 3:

NCI-H929 whole cell extract at 30 µg

Predicted band size: 82 kDa

false

• WB

Supplier Data

Western blot - Anti-SP3 antibody - N-terminal (AB227856)

7.5% SDS-PAGE gel.

All lanes:

Western blot - Anti-SP3 antibody - N-terminal (ab227856) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human epithelial cell line from cervix adenocarcinoma) cell extract at 30 µg

Lane 2:

SP3 knockout HeLa (human epithelial cell line from cervix adenocarcinoma) cell extract at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Predicted band size: 82 kDa

true

• WB

Lab

Western blot - Anti-SP3 antibody - N-terminal (AB227856)

All lanes:

Western blot - Anti-SP3 antibody - N-terminal (ab227856) at 1/1000 dilution

Lane 1:

WT HeLa cell extract at 30 µg

Lane 2:

SP3 Knockout HeLa cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Predicted band size: 82 kDa

false

• WB

Supplier Data

Western blot - Anti-SP3 antibody - N-terminal (AB227856)

7.5 % SDS-PAGE gel.

All lanes:

Western blot - Anti-SP3 antibody - N-terminal (ab227856) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell extract at 30 µg

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) nuclear extract at 30 µg

Predicted band size: 82 kDa

false