Anti-Senataxin antibody [EPR24137-74] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Senataxin antibody [EPR24137-74] - BSA and Azide free (AB300440)

This data was developed using ab300439, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Senataxin with ab300439 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-Senataxin antibody [EPR24137-74] - BSA and Azide free (AB300440)

This data was developed using ab300439, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer and concentration : 5% NFDM/TBST

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17562789).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-Senataxin antibody [EPR24137-74] (<a href='/en-us/products/primary-antibodies/senataxin-antibody-epr24137-74-ab300439'>ab300439</a>) at 1/1000 dilution

Lane 1:

HeLa transfected with scrambled siRNA control, whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting SETX, whole cell lysate at 20 µg

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 1/100000 dilution

Predicted band size: 302 kDa

Observed band size: 300 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Senataxin antibody [EPR24137-74] - BSA and Azide free (AB300440)

This data was developed using ab300439, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer and concentration : 5% NFDM/TBST

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17562789).

The lysate was freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-Senataxin antibody [EPR24137-74] (<a href='/en-us/products/primary-antibodies/senataxin-antibody-epr24137-74-ab300439'>ab300439</a>) at 1/1000 dilution

All lanes:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

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 1/100000 dilution

Predicted band size: 302 kDa

Observed band size: 300 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Senataxin antibody [EPR24137-74] - BSA and Azide free (AB300440)

This data was developed using ab300439, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer and concentration : 5% NFDM/TBST

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17562789).

The bands beneath the target band are likely to be degraded target fragments.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-Senataxin antibody [EPR24137-74] (<a href='/en-us/products/primary-antibodies/senataxin-antibody-epr24137-74-ab300439'>ab300439</a>) at 1/1000 dilution

Lane 1:

SK-N-BE(2) (human neuroblastoma neuroblast), whole cell lysate at 20 µg

Lane 2:

IMR-32 (human neuroblastoma neuroblast), whole cell lysate at 20 µg

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 1/100000 dilution

Predicted band size: 302 kDa

Observed band size: 300 kDa

false

Exposure time: 180s

• WB

Collaborator

Western blot - Anti-Senataxin antibody [EPR24137-74] - BSA and Azide free (AB300440)

ab300439 was shown to react with SETX in wild-type RKO cells in Western blot with loss of signal observed in SETX knockout cell line ab324539. Wild-type RKO and SETX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab300439 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was 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.

All lanes:

Western blot - Anti-Senataxin antibody [EPR24137-74] (<a href='/en-us/products/primary-antibodies/senataxin-antibody-epr24137-74-ab300439'>ab300439</a>) at 1/500 dilution

Lane 1:

Wild-type RKO lysate at 50 µg

Lane 2:

Western blot - Human SETX knockout RKO cell line (<a href='/en-us/products/cell-lines/human-setx-knockout-rko-cell-line-ab324539'>ab324539</a>) at 50 µg

Predicted band size: 302 kDa

Observed band size: 300 kDa

false