Anti-SKP2 antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SKP2 antibody (AB68455)

Immunofluorescence analysis of SKP2 using Anti-SKP2 antibody. SKP2 was detected in a immunocytochemical section of SiHa cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 minutes. The cells were blocked with 10% goat serum. The cells were then incubated with 5 μg/mL ab68455 and Mouse anti-Tubulin Alpha antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG and Cy3 Conjugated Goat Anti-Mouse IgG were used as secondary antibodies at 1 : 500 dilution and incubated for 30 minutes at 37°C. Visualized using a fluorescence microscope and filter sets appropriate for the label used.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-SKP2 antibody (AB68455)

Flow cytometric analysis of 4% paraformaldehyde fixed and permeabilized HepG2 cells. The cells were blocked with 10% normal goat serum, then incubated with ab68455 (1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was Rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

• WB

Supplier Data

Western blot - Anti-SKP2 antibody (AB68455)

Blocking buffer : 5% non-fat milk/TBS

All lanes:

Western blot - Anti-SKP2 antibody (ab68455) at 0.5 µg/mL

Lane 1:

Human 293T whole cell lysates at 30 µg

Lane 2:

Human Jurkat whole cell lysates at 30 µg

Lane 3:

Human HepG2 whole cell lysates at 30 µg

Lane 4:

Human MCF-7 whole cell lysates at 30 µg

Secondary

All lanes:

Goat anti-rabbit IgG-HRP secondary antibody at 1/5000 dilution

Observed band size: 48 kDa

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• WB

Unknown

Western blot - Anti-SKP2 antibody (AB68455)

The predicted molecular weight of SKP2 is 48 kDa (SwissProt), however we expect to observe a banding pattern at 53 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-SKP2 antibody (ab68455) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 3:

Human placenta tissue lysate - total protein (<a href='/en-us/products/unavailable/human-placenta-tissue-lysate-total-protein-ab29745'>ab29745</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 47 kDa

Observed band size: 29 kDa,38 kDa,53 kDa

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Exposure time: 1min

• WB

CiteAb

Western blot - Anti-SKP2 antibody (AB68455)

SKP2 western blot using anti-SKP2 antibody ab68455. Publication image and figure legend from Ma, X., Zhao, J., et al., 2017, Oncotarget, PubMed 29137415.

ab68455 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 ab68455 please see the product overview.

UBE2L3 interacted with SCF (Skp2) complex and promoted p27kip1 ubiquitination and proteasomal degradation(A) Knockdown of UBE2L3 in A549 and NCI-H1299 cells promoted the protein level of p27kip1. (B) Overexpression of UBE2L3 in NCI-H460 and HCC827 cells decreased the protein level of p27kip1. (C) The association between UBE2L3 and SCF (Skp2) complex and p27kip1 was confirmed by immunoprecipitation and immunoblot assay. (D) Overexpression of UBE2L3 promoted the ubiquitination and proteasomal degradation of p27kip1. (E, F) The half-life of p27kip1 was evaluated by CHX assay after UBE2L3 overexpression.

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