Anti-USP13 抗体 [EPR4348]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-USP13 antibody [EPR4348] (AB109264)

ICC/IF image of ab109264 stained MCF-7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab109264 at 10μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-USP13 antibody [EPR4348] (AB109264)

Overlay histogram showing SHSY-5Y cells stained with ab109264 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109264 1/9400 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-USP13 antibody [EPR4348] (AB109264)

Immunocytochemistry/Immunofluorescence analysis of Neuro-2a (Mouse neuroblastoma cell line) labeling USP13 with Purified ab109264 at 1/500 dilution (5 μg/ml). Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• WB

Lab

Western blot - Anti-USP13 antibody [EPR4348] (AB109264)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : USP13 knockout HAP1 cell lysate (20 μg)
Lane 3 : A375 cell lysate (20 μg)
Lane 4 : SH-SY5Y cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab109264 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109264 was shown to recognize USP13 when USP13 knockout samples were used, along with additional cross-reactive bands. Wild-type and USP13 knockout samples were subjected to SDS-PAGE. ab109264 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-USP13 antibody [EPR4348] (ab109264)

Predicted band size: 97 kDa

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• WB

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Western blot - Anti-USP13 antibody [EPR4348] (AB109264)

All lanes:

Western blot - Anti-USP13 antibody [EPR4348] (ab109264) at 1/1000 dilution

Lane 1:

SH SY5Y cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

A375 cell lysate at 10 µg

Predicted band size: 97 kDa

Observed band size: 97 kDa

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• WB

CiteAb

Western blot - Anti-USP13 antibody [EPR4348] (AB109264)

USP13 western blot using anti-USP13 antibody [EPR4348] ab109264. Publication image and figure legend from Liao, Y., Guo, Z., et al., 2019, J Exp Clin Cancer Res, PubMed 30975171.

ab109264 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 ab109264 please see the product overview.

Spautin-1 induces G0/G1 phase arrest via down-regulating Cyclin D1 in PCa cells. a Fluorescence-activated cell sorting analysis (FACS) was performed to analyze cell cycle distributions of PCa cells exposed to Spautin-1 or SKP2-C25 for 24 h. A summary of cell cycle distributions was shown from three independent experiments. b Western blot analysis was performed to detect the expression of CDK4, CDK2, Cyclin D1, P15 and P21 in PCa cells exposed to various doses of Spautin-1 (0, 5, 10, 20 μM) for 24 h (left), or Spautin-1 (10 μM) at various lengths of time (Right). c Western blot analysis was performed to detect the expression of CDK4, CDK2, Cyclin D1, USP10 and USP13 in PCa cells exposed to USP10 siRNA or USP13 siRNA for 48 h. d FACS was performed to analyze cell cycle distributions of PCa cells exposed to USP10 siRNA or USP13 siRNA for 48 h. e 22Rv1 and PC3 cells were transfected with HA-Cyclin D1 and/or FLAG-CDK2 for 48 h. Cell viability analysis was performed on the above cells exposed to Spautin-1 for 24 h. #p<0.05. f Western blot of CDK2 and Cyclin D1 to verified the overexpression in PC3 cells. Sp-1 : Spautin-1

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