Anti-Drebrin antibody [mAbcam60932]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Drebrin antibody [mAbcam60932] (AB60932)

Overlay histogram showing SHSY-5Y cells stained with ab60932 (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 (ab60932, 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 IgG1 [ICIGG1] (ab91353, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. 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.

• WB

Lab

Western blot - Anti-Drebrin antibody [mAbcam60932] (AB60932)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : Drebrin knockout HAP1 whole cell lysate (20 μg)
Lane 3 : SH-SY5Y whole cell lysate (20 μg)

Lanes 1 - 3 : Merged signal (red and green). Green - ab60932 observed at 120 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab60932 was shown to specifically react with Drebrin in wild-type cells as signal was lost in Drebrin knockout cells. Wild-type and Drebrin knockout samples were subjected to SDS-PAGE. ab60932 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 5 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Drebrin antibody [mAbcam60932] (ab60932)

Predicted band size: 71 kDa

false

• WB

Unknown

Western blot - Anti-Drebrin antibody [mAbcam60932] (AB60932)

ab60932 detects a band around 110-kDa. While this differs to its predicted molecular weight of 71-kDa, it is consistent with what has been described in the literature(PMID : 10604653).

All lanes:

Western blot - Anti-Drebrin antibody [mAbcam60932] (ab60932) at 10 µg/mL

Lane 1:

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

Lane 2:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 71 kDa

Observed band size: 10 kDa,110 kDa,65 kDa

true

Exposure time: 8min

• WB

Unknown

Western blot - Anti-Drebrin antibody [mAbcam60932] (AB60932)

All lanes:

Western blot - Anti-Drebrin antibody [mAbcam60932] (ab60932) at 10 µg/mL

Lane 1:

Human Cortex Neuronal cell lysate at 10 µg

Lane 2:

Rat Cortex Neuronal cell lysate at 10 µg

Lane 3:

Rat Hippocampus Tissue Lysate at 10 µg

Lane 4:

Human skin tumor tissue lysate (melanoma) - total protein (<a href='/en-us/products/unavailable/human-skin-tumor-tissue-lysate-melanoma-total-protein-ab29339'>ab29339</a>) at 10 µg

Lane 5:

Malme 3M (Human melanoma cells) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 71 kDa

Observed band size: 100 kDa,120 kDa,200 kDa,45 kDa,50 kDa,60 kDa

false