Anti-DDX5 抗体

• WB

Lab

Western blot - Anti-DDX5 antibody (AB21696)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : DDX5 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : MCF7 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab21696 observed at 69 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab21696 was shown to specifically react with DDX5 in wild-type HAP1 cells as signal was lost in DDX5 knockout cells. Wild-type and DDX5 knockout samples were subjected to SDS-PAGE. ab21696 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-DDX5 antibody (ab21696)

Predicted band size: 69 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DDX5 antibody (AB21696)

ab21696 staining DDX5 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab21696 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Project

Western blot - Anti-DDX5 antibody (AB21696)

ab21696 recognizes a clear, strong band at ~ 69kDa corresponding to DDX5. A number of other, non specific bands are weakly recognized by the antibody. All bands are quenched by the addition of the blocking peptide.

All lanes:

Western blot - Anti-DDX5 antibody (ab21696) at 1 µg/mL

Lane 1:

HeLa nuclear lysate at 20 µg

Lane 2:

HeLa nuclear lysate at 20 µg with Human DDX5 peptide (ab22412)

Secondary

All lanes:

Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

Predicted band size: 69 kDa

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• WB

Project

Western blot - Anti-DDX5 antibody (AB21696)

All lanes:

Western blot - Anti-DDX5 antibody (ab21696) at 1 µg/mL

Lane 1:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 2:

Testis (Mouse) Tissue Lysate - normal tissue at 10 µg

Lane 3:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 69 kDa

Observed band size: 170 kDa,70 kDa

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