Anti-MYO5B antibody [EPR28275-55]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-MYO5B antibody [EPR28275-55] (AB313795)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Mouse primary neuron cells labelling MYO5B with ab313795 at 1/150 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081 secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). The nuclear counter stain is DAPI (blue).

The negative controls are as follows :
-ve control 1 : primary ab313795 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary at 1/1000 dilution.
-ve control 2 : primary ab11267 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MYO5B antibody [EPR28275-55] (AB313795)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neuron cells labelling MYO5B with ab313795 at 1/50 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MYO5B antibody [EPR28275-55] (AB313795)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling MYO5B with ab313795 at 1/150 dilution (1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-MYO5B antibody [EPR28275-55] (AB313795)

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/1000000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MYO5B antibody [EPR28275-55] (ab313795) at 1/1000 dilution

Lane 1:

HaCaT (human skin keratinocyte) whole cell lysate at 20 µg

Lane 2:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 213 kDa,37 kDa

Observed band size: 213 kDa,37 kDa

false

Exposure time: 60s

• WB

Lab

Western blot - Anti-MYO5B antibody [EPR28275-55] (AB313795)

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/1000000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lanes 1 - 2:

Western blot - Anti-MYO5B antibody [EPR28275-55] (ab313795) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-MYO5B antibody [EPR28275-55] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/myo5b-antibody-epr28275-55-bsa-and-azide-free-ab313796'>ab313796</a>)

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 213 kDa,37 kDa

Observed band size: 213 kDa,37 kDa

false

Exposure time: 120s

• WB

Lab

Western blot - Anti-MYO5B antibody [EPR28275-55] (AB313795)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Rat primary neuron cells labelling MYO5B with ab313795 at 1/150 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining in rat primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). The nuclear counter stain is DAPI (blue)

The negative controls are as follows : -ve control 1 : primary ab313795 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution. -ve control 2 : primary ab11267 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary at 1/1000 dilution.

• Dot

Lab

Dot Blot - Anti-MYO5B antibody [EPR28275-55] (AB313795)

Dot blot analysis of MYO5B using ab313795 at 1 : 1000 (0.527 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. This antibody does not cross-react with human MYO5A, but has a weak cross-reactivity with human MYO5C. Blocking and diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Dot Blot - Anti-MYO5B antibody [EPR28275-55] (ab313795) at 1/1000 dilution

Lane 1:

MYO5B peptide

Lane 2:

MYO5A peptide

Lane 3:

MYO5C peptide

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s