Anti-HOXB13 antibody [mAbcam53931]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-HOXB13 antibody [mAbcam53931] (AB53931)

Overlay histogram showing HeLa cells stained with ab53931 (red line). The cells were fixed with 80% methanol (5 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 (ab53931, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HOXB13 antibody [mAbcam53931] (AB53931)

ab53931 staining HOXB13 in HepG2 cells. The cells were fixed with 4% paraformaldehyde (10 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 ab53931 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

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-HOXB13 antibody [mAbcam53931] (AB53931)

ab53931 recognizes the tagged HOXB13 recombinant protein which has an expected molecular weight of 57 kDa.

All lanes:

Western blot - Anti-HOXB13 antibody [mAbcam53931] (ab53931) at 5 µg/mL

All lanes:

HOXB13 Recombinant protein at 0.1 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 30 kDa

Observed band size: 57 kDa

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