重组Anti-SAMD9抗体[EPR13603]

• WB

Lab

Western blot - Anti-SAMD9 antibody [EPR13603] (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] (ab180575) 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='/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='/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

Supplier Data

Western blot - Anti-SAMD9 antibody [EPR13603] (AB180575)

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

All lanes:

Western blot - Anti-SAMD9 antibody [EPR13603] (ab180575) at 1/10000 dilution

Lane 1:

A431 cell lysate at 20 µg

Lane 2:

MCF-7 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-rabbit HRP at 1/1000 dilution

Predicted band size: 184 kDa

false

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SAMD9 antibody [EPR13603] (AB180575)

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).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SAMD9 antibody [EPR13603] (AB180575)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixedA431 cells labeling SAMD9 with ab180575 (unpurified) at 1/50 dilution, or Rabbit monoclonal IgG Isotype control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SAMD9 antibody [EPR13603] (AB180575)

Immunofluorescence analysis of 4% paraformaldehyde-A431 cells labeling SAMD9 with ab180575 (unpurified) (green) at 1/100 dilution. Dapi counterstain (blue). Goat anti rabbit IgG (Dylight 488) at a dilution of 1/200 was used as a secondary antibody.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SAMD9 antibody [EPR13603] (AB180575)

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.

• WB

Lab

Western blot - Anti-SAMD9 antibody [EPR13603] (AB180575)

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] (ab180575) 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='/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='/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] (AB180575)

All lanes:

Western blot - Anti-SAMD9 antibody [EPR13603] (ab180575) at 1/1000 dilution

All lanes:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 184 kDa

Observed band size: 184 kDa

false