Anti-Myoferlin antibody [EPR18887] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Myoferlin antibody [EPR18887] - BSA and Azide free (AB271928)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Myoferlin with ab178386 at 1/400 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178386).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Myoferlin antibody [EPR18887] - BSA and Azide free (AB271928)

Immunofluorescent analysis of 100% methanol-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Myoferlin with ab178386 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab178386 at 1/100 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178386).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Myoferlin antibody [EPR18887] - BSA and Azide free (AB271928)

Immunofluorescent analysis of 100% methanol-fixed C2C12 (mouse myoblast cell line) cells labeling Myoferlin with ab178386 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on C2C12 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab178386 at 1/100 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178386).

• WB

Lab

Western blot - Anti-Myoferlin antibody [EPR18887] - BSA and Azide free (AB271928)

This data was developed using the same antibody clone in a different buffer formulation (ab178386).

Lanes 1-4 : Merged signal (red and green). Green - ab178386 observed at 250,180 kDa. Red - loading control ab7291 observed at 50 kDa.

ab178386 Anti-Myoferlin antibody [EPR18887] was shown to specifically react with Myoferlin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265782 (knockout cell lysate ab257547) was used. Wild-type and Myoferlin knockout samples were subjected to SDS-PAGE. ab178386 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-Myoferlin antibody [EPR18887] (<a href='/en-us/products/primary-antibodies/myoferlin-antibody-epr18887-ab178386'>ab178386</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MYOF knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MYOF (Myoferlin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-myof-myoferlin-knockout-hela-cell-line-ab265782'>ab265782</a>)

Lane 3:

MDA-MB-231 cell lysate at 20 µg

Lane 4:

SK-BR-3 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: 235 kDa

Observed band size: 180 kDa,250 kDa

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