Anti-SLFN12 antibody [EPR20904-32]

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-SLFN12 antibody [EPR20904-32] (AB234418)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen human placenta tissue labeling SLFN12 with ab234418 at 1/100 dilution (green), followed by Donkey anti-Rabbit IgG (H+L) secondary antibody, Alexa Fluor^® 555 secondary at a 1/100 dilution. Positive staining in syncytiotrophoblast of human placenta is observed. Counterstained with DAPI (blue). Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Donkey anti-Rabbit IgG (H+L) secondary antibody, Alexa Fluor^® 555 used at a 1/100 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).

The image is kindly provided by Dr Xiaodong Wang.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SLFN12 antibody [EPR20904-32] (AB234418)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling SLFN12 with ab234418 at 1/60 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

• WB

Unknown

Western blot - Anti-SLFN12 antibody [EPR20904-32] (AB234418)

Blocking/Dilution buffer : 5% Defatted milk.

The image is kindly provided by Dr Xiaodong Wang.

ab234418 was shown to specifically react with SLFN12 in wild-type Hela cells as signal was lost in SLFN12 knockout cells. Wild-type and SLFN12 knockout samples were subjected to SDS-PAGE. ab234418 and anti-GAPDH antibody were incubated 8 hours at 4 °C at 1/500 dilution and 1/2000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (whole molecule), Peroxidase conjugated secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument using the ECL technique.

All lanes:

Western blot - Anti-SLFN12 antibody [EPR20904-32] (ab234418) at 1/500 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) treated with DMSO for 12 hours, whole cell lysate at 30 µg

Lane 2:

Wild-type HeLa treated with 17β-estradiol (5 μM) for 12 hours, whole cell lysate at 30 µg

Lane 3:

SLFN12 knockout HeLa treated with DMSO for 12 hours, whole cell lysate at 30 µg

Lane 4:

SLFN12 knockout HeLa treated with 17β-estradiol (5 μM) for 12 hours, whole cell lysate at 30 µg

Secondary

All lanes:

Anti-Rabbit IgG (whole molecule)–Peroxidase antibody produced in goat at 1/2000 dilution

Predicted band size: 66 kDa

Observed band size: 67 kDa

false

Exposure time: 5s

• WB

Collaborator

Western blot - Anti-SLFN12 antibody [EPR20904-32] (AB234418)

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

All lanes:

Western blot - Anti-SLFN12 antibody [EPR20904-32] (ab234418) at 1/500 dilution

Lane 1:

HEK-293 (human embryonic kidney) transfected with an empty vector (vector control) containing HA- tag® whole cell lysates at 15 µg

Lane 2:

HEK-293 (human embryonic kidney) transfected with SLFN12 (Q8IYM2) (WT) expression vector containing HA-tag®, whole cell lysate at 15 µg

Secondary

All lanes:

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

Predicted band size: 66 kDa

Observed band size: 67 kDa

true

Exposure time: 10s

SLFN12 plasmid was kindly provided by an anonymous collaborator.