Anti-MYL9 antibody [EPR13012(2)]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling MYL9 with ab191393 at 1/700 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin. Inset : Negative control : using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling MYL9 with ab191393 at 1/700 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin. Inset : Negative control : using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

Lanes 1-4 : Merged signal (red and green). Green - ab191393 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.

ab191393 Anti-MYL9 antibody [EPR13012(2)] was shown to specifically react with MYL9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266036 (knockout cell lysate ab256999) was used. Wild-type and MYL9 knockout samples were subjected to SDS-PAGE. ab191393 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MYL9 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-myl9-knockout-hela-cell-lysate-ab256999'>ab256999</a>) at 20 µg

Lane 3:

Human colon cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 20 kDa

Observed band size: 20 kDa,37 kDa

false

• WB

Supplier Data

Western blot - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

All lanes:

Western blot - Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/1000 dilution

Lane 1:

Human stomach lysate at 20 µg

Lane 2:

Human colon cancer lysate at 20 µg

Lane 3:

Human uterus lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 20 kDa

false

• WB

Supplier Data

Western blot - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

All lanes:

Western blot - Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/1000 dilution

Lane 1:

C6 cell lysate at 10 µg

Lane 2:

PC12 cell lysate at 10 µg

Lane 3:

NIH 3T3 cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 20 kDa

false

• WB

Supplier Data

Western blot - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

The identity of the higher MW bands are unknown.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-MYL9 antibody [EPR13012 (2)] (ab191393) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-MYL9 (pS20) + MYL12A (pS19) + MYL12B (pS20) antibody [EPR27203-65] (<a href='/en-us/products/primary-antibodies/myl9-ps20-myl12a-ps19-myl12b-ps20-antibody-epr27203-65-ab316750'>ab316750</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate (untreated membrane) at 45 µg with 5% NFDM/TBST

Lane 2:

HeLa treated with 100ng/ml Calyculin A for 15 minutes, whole cell lysate (untreated membrane) at 45 µg with 5% NFDM/TBST

Lane 3:

Untreated HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate (alkaline phosphatase treated membrane) at 45 µg with 5% NFDM/TBST

Lane 4:

HeLa treated with 100ng/ml Calyculin A for 15 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 45 µg with 5% NFDM/TBST

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

Observed band size: 20 kDa,15 kDa

false

Exposure time: 70s

• IP

Supplier Data

Immunoprecipitation - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

Western blot analysis of immunoprecipitation pellet from Human stomach lysate immunoprecipitated using ab191393 at 1/40 dilution (lane 1) or PBS control (lane 2).
Secondary : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

All lanes:

Immunoprecipitation - Anti-MYL9 antibody [EPR13012(2)] (ab191393)

Predicted band size: 20 kDa

false

• WB

Supplier Data

Western blot - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

All lanes:

Western blot - Anti-MYL9 antibody [EPR13012(2)] (ab191393) at 1/5000 dilution

All lanes:

rat stomach lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 20 kDa

false

• WB

Supplier Data

Western blot - Anti-MYL9 antibody [EPR13012(2)] (AB191393)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

In Western blot, Anti-MYL9 antibody [EPR13012 (2)] (ab191393) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-MYL9 (pS20) + MYL12A (pS19) + MYL12B (pS20) antibody [EPR27203-65] (<a href='/en-us/products/primary-antibodies/myl9-ps20-myl12a-ps19-myl12b-ps20-antibody-epr27203-65-ab316750'>ab316750</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with human MYL9 expression vector containing a myc-His-tag® then treated with 100/ml calyculin for 15 minutes, whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) cells transfected with human MYL9 (S20A mutant) expression vector containing a myc-His-tag® then treated with 100/ml calyculin for 15 minutes, whole cell lysate at 20 µg with 5% NFDM/TBST

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

Observed band size: 25 kDa,36 kDa

false

Exposure time: 180s