Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Myocilin with ab318197 at 1/500 dilution (0.1ug) / Magenta compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Myocilin with ab318197 at 1/500 (1.016 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse retina cells labelling Myocilin with ab318197 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype contro.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary retina cells labelling Myocilin with ab318197 at 1/500 (1.016 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in rat primary retina (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.
The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab318197 at 1/500 (1.016 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2 ug/mL dilution.
-ve control 2 : ab11267 1/500 4ug/ml dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat retina cells labelling Myocilin with ab318197 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype contro.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary retina cells labelling Myocilin with ab318197 at 1/500 (1.016 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary retina (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.
The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab318197 at 1/500 (1.016 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2 ug/mL dilution.
-ve control 2 : ab11267 1/500 4ug/ml dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• WB

Supplier Data

Western blot - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Low expression : Human spleen and liver (PMID : 10320784).

The bands beneath the target band (20~37 kDa) are likely to be degraded target fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Myocilin antibody [EPR28799-15] (<a href='/en-us/products/primary-antibodies/myocilin-antibody-epr28799-15-ab318197'>ab318197</a>) at 1/1000 dilution

Lane 1:

Human eyeball tissue lysate at 20 µg

Lane 2:

Human testis tissue lysate at 20 µg

Lane 3:

Human spleen tissue lysate at 20 µg

Lane 4:

Human liver 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 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

The identity of the higher MW band at approximately 150kDa (Lane 3) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-3 : 92 seconds; Lane 4-6 : 180 seconds

All lanes:

Western blot - Anti-Myocilin antibody [EPR28799-15] (<a href='/en-us/products/primary-antibodies/myocilin-antibody-epr28799-15-ab318197'>ab318197</a>) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole fresh cell lysate at 20 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole fresh cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole fresh cell lysate at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

C2C12 (mouse myoblast) 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 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Myocilin antibody [EPR28799-15] - BSA and Azide free (AB318198)

This data was developed using ab318197, the same antibody clone in a different buffer formulation.

Low expression : Mouse and rat liver (PMID : 10320784).

The identity of the lower MW band at approximately 37kDa (Lane 3) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Myocilin antibody [EPR28799-15] (<a href='/en-us/products/primary-antibodies/myocilin-antibody-epr28799-15-ab318197'>ab318197</a>) at 1/1000 dilution

Lane 1:

Mouse retina tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Rat retina tissue lysate at 20 µg

Lane 5:

Rat testis tissue lysate at 20 µg

Lane 6:

Rat liver 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 1/100000 dilution

Observed band size: 55 kDa,36 kDa

false

Exposure time: 92s