Anti-Spastin抗体[Sp 6C6] (ab77144)

ab77144 was shown to react with SPAST in wild-type HEK293T cells in Western blot with loss of signal observed in SPAST knockout cell line ab267238 (SPAST knockout cell lysate ab258698). Wild-type HEK293T and SPAST knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab77144 overnight at 4 °C at a 1/500 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2μg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

Immunoprecipitation of SPAST in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 1µ of ab77144 pre-coupled to Protein A beads. Samples were washed and processed for western blot with ab77144 at 1/500.

This data was kindly provided by the YCharOS Inc., an open science company with the mission of characterizing every commercially available antibody reagent. Abcam are working with YCharOS to support their mission of antibody characterisation using knock out cell lines.

Lanes 1-3: Merged signal (red and green). Green - ab77144 observed at 52 kDa. Red - loading control ab181602 observed at 36 kDa.

 ab77144 Anti-Spastin antibody [Sp 6C6] was shown to specifically react with Spastin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267238 (knockout cell lysate ab258698) was used. Wild-type and Spastin knockout samples were subjected to SDS-PAGE. ab77144 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

9% SDS-PAGE.

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