Anti-SPOP antibody

• WB

Lab

Western blot - Anti-SPOP antibody (AB137537)

Western blot : Anti-SPOP antibody (ab137537) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab137537 was shown to bind specifically to SPOP. A band was observed at 42 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in SPOP knockout cell line ab277845. To generate this image, wild-type and SPOP knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

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Western blot - Anti-SPOP antibody (ab137537) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

SPOP knockout HEK-293 cell lysate at 20 µg

Lane 3:

Recombinant Human SPOP protein <a href='/en-us/products/proteins-peptides/recombinant-human-spop-protein-ab105599'>ab105599</a> cell lysate at 0.1 µg

Secondary

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Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 42 kDa

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• WB

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Western blot - Anti-SPOP antibody (AB137537)

10% SDS PAGE

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Western blot - Anti-SPOP antibody (ab137537) at 1/2000 dilution

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Mouse brain lysate at 50 µg

Secondary

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HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 42 kDa

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• WB

Supplier Data

Western blot - Anti-SPOP antibody (AB137537)

10% SDS-PAGE

All lanes:

Western blot - Anti-SPOP antibody (ab137537) at 1/5000 dilution

Lane 1:

Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 30 µg

Lane 2:

Transfected HEK-293T whole cell lysate at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 42 kDa

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• WB

CiteAb

Western blot - Anti-SPOP antibody (AB137537)

SPOP western blot using anti-SPOP antibody ab137537. Publication image and figure legend from Zhang, P., Zhang, Y., et al., 2015, Oncotarget, PubMed 26595804.

ab137537 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab137537 please see the product overview.

ASPP1/2 are required for proper mitotic progressiona. Depletion of ASPP1/2 by siRNAs in HeLa cells. HeLa cells were transfected with the indicated siRNAs. After 48 hr, cell lysates were prepared for WB analyses with their indicated antibodies. b. ASPP1/2 co-depletion causes G2/M arrest. The cell-cycle distributions of HeLa cells transfected with indicated siRNAs for 48 hr were determined by flow cytometry. Error bars, SEM. *p<0.01 from triplicates. c. ASPP1/2 co-depletion increases the mitotic index in HeLa cells. HeLa cells were transfected with control or ASPP1/2 siRNAs as indicated. After 48 hr, cells were fixed and stained for the p-H3 (Ser10) antibody. Quantification of cells with anti-p-H3 (Ser10) staining is shown at the right (n= number of counted cells). d. Mitotic stages were quantified by DNA and spindle morphology in the mitotic population of control or ASPP1/2 co-depleted HeLa cells. e. ASPP1/2 co-depletion increases the number of mitotic cells with misaligned chromosomes. HeLa cells were transfected with control or ASPP1/2 siRNAs. After 48 hr, cells were fixed and stained with the anti-β-tubulin (green) antibody and DAPI (blue). White arrows indicate misaligned chromosomes. Scale bar = 10μm. Quantification of cells with misaligned chromosomes is shown on the right. f. ASPP1/2 depletions lead to increases in cell death. HeLa cells were transfected with control or ASPP1/2 siRNAs. After 72hr, the cell death was measured by flow cytometry using the propidium iodide staining assay.

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