Anti-SAMD9 antibody [EPR13603] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SAMD9 antibody [EPR13603] - BSA and Azide free (AB250226)

Intracellular Flow Cytometry analysis of MCF-7 (Human breast adenocarcinoma epithelial cell) cells labeling SAMD9 with purified ab180575 at 1/30 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180575)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SAMD9 antibody [EPR13603] - BSA and Azide free (AB250226)

Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling SAMD9 with purified ab180575 at 1 : 50 dilution (5.4 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with none. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (anti samd9 antibody epr13603 immunocytochemistry a431 human)

• WB

Lab

Western blot - Anti-SAMD9 antibody [EPR13603] - BSA and Azide free (AB250226)

This data was developed using ab180575, the same antibody clone in a different buffer formulation.

Lanes 1-3 : Merged signal (red and green). Green - ab180575 observed at 184 kDa. Red - loading control ab8245 observed at 36 kDa.

ab180575 Anti-SAMD9 antibody [EPR13603] was shown to specifically react with SAMD9 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267039 (knockout cell lysate ab257657) was used. Wild-type and SAMD9 knockout samples were subjected to SDS-PAGE. ab180575 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-SAMD9 antibody [EPR13603] (<a href='/en-us/products/primary-antibodies/samd9-antibody-epr13603-ab180575'>ab180575</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SAMD9 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human SAMD9 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-samd9-knockout-a549-cell-line-ab267039'>ab267039</a>)

Lane 3:

A431 cell lysate at 20 µg

Secondary

All lanes:

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 1/10000 dilution

Predicted band size: 184 kDa

Observed band size: 184 kDa

false

• WB

Lab

Western blot - Anti-SAMD9 antibody [EPR13603] - BSA and Azide free (AB250226)

This data was developed using the same antibody clone in a different buffer formulation (ab180575).

Lanes 1-4 : Merged signal (red and green). Green - ab180575 observed at 184 kDa. Red - loading control ab8245 observed at 36 kDa.

ab180575 Anti-SAMD9 antibody [EPR13603] was shown to specifically react with SAMD9 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267038 (knockout cell lysate ab257656) was used. Wild-type and SAMD9 knockout samples were subjected to SDS-PAGE. ab180575 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-SAMD9 antibody [EPR13603] (<a href='/en-us/products/primary-antibodies/samd9-antibody-epr13603-ab180575'>ab180575</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SAMD9 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human SAMD9 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-samd9-knockout-a549-cell-line-ab267038'>ab267038</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Human skeletal muscle cell lysate at 20 µg

Secondary

All lanes:

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 1/10000 dilution

Predicted band size: 184 kDa

Observed band size: 184 kDa

false