Anti-Musashi 1 / Msi1 antibody [Y2709]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Musashi 1 / Msi1 antibody [Y2709] (AB129819)

ab129819 staining Musashi 1 / Msi1 in HAP1-MSI1 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween 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 ab129819 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 red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• WB

Lab

Western blot - Anti-Musashi 1 / Msi1 antibody [Y2709] (AB129819)

Lanes 1 - 3 : Merged signal (red and green). Green - ab129819 observed at 39 kDa. Red - loading control, ab52866, observed at 50 kDa.

ab129819 was shown to specifically react with Musashi 1 / Msi1 in wild-type HAP1 cells as signal was lost in MSI1 knockout cells. Wild-type and MSI1 knockout samples were subjected to SDS-PAGE. ab129819 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 5 ug/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-Musashi 1 / Msi1 antibody [Y2709] (ab129819) at 5 µg/mL

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

MSI1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

SH-SY5Y whole cell lysate at 20 µg

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

• WB

Project11284****

Western blot - Anti-Musashi 1 / Msi1 antibody [Y2709] (AB129819)

All lanes:

Western blot - Anti-Musashi 1 / Msi1 antibody [Y2709] (ab129819) at 5 µg/mL

Lane 1:

Brain (Human) Membrane Lysate - fetal normal tissue (<a href='/en-us/products/unavailable/brain-human-membrane-lysate-fetal-normal-tissue-ab29457'>ab29457</a>) at 20 µg

Lane 2:

E14 Rat Embryo Brain Tissue Lysate at 20 µg

Lane 3:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 20 µg

Lane 4:

46C (Mouse neural progenitor, selected for Sox1 expression cell line) Whole Cell Lysate (<a href='/en-us/products/unavailable/46c-mouse-neural-progenitor-selected-for-sox1-expression-cell-line-whole-cell-lysate-ab27205'>ab27205</a>) at 20 µg

Secondary

All lanes:

Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/10000 dilution

Predicted band size: 39 kDa

Observed band size: 40 kDa

true

Exposure time: 12min