Anti-DKK1 antibody [EPR4759]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (AB109416)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue using 1/100 unpurified ab109416.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKK1 antibody [EPR4759] (AB109416)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling DKK1 with Purified ab109416 at 1 : 800 dilution (1.04 µg/ml). Heat mediated antigen retrieval was performed using using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

• WB

Lab

Western blot - Anti-DKK1 antibody [EPR4759] (AB109416)

False colour image of Western blot : Anti-DKK1 antibody [EPR4759] staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-DKK1 antibody [EPR4759] (ab109416) at 1/2000 dilution

Lane 1:

Caco-2 cell lysate at 20 µg

Lane 2:

PC-3 cell lysate at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

MOLT4 cell lysate at 20 µg

Lane 5:

HEK-293 cell lysate at 20 µg

Predicted band size: 29 kDa

Observed band size: 30-45 kDa

false

• WB

Lab

Western blot - Anti-DKK1 antibody [EPR4759] (AB109416)

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

HEK-293 and 293T cell are weak expressed cell lines for DKK1 protein (PMID : 18502762, PMID : 15378020).

All lanes:

Western blot - Anti-DKK1 antibody [EPR4759] (ab109416) at 1/10000 dilution

Lane 1:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HEK-293 (Human embryonic kidney 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 1/20000 dilution

Predicted band size: 29 kDa

Observed band size: 29-40 kDa

false

Exposure time: 180s

• WB

Unknown

Western blot - Anti-DKK1 antibody [EPR4759] (AB109416)

Blocking and diluting buffer : 5% NFDM/TBST.

The smear bands detected by ab109416 are similar to those reported by the papers (PMID : 18546262, 28090290, 24008318), which should be due to the glycosylation of DKK1.

All lanes:

Western blot - Anti-DKK1 antibody [EPR4759] (ab109416) at 1/2000 dilution

Lane 1:

A549 (Human lung carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

NCI-H460 (Human Large Cell Lung Cancer epithelial cell) whole cell lysates at 20 µg

Lane 4:

DU145 (Human prostate carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 5:

SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysates 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 1/20000 dilution

Predicted band size: 29 kDa

false

• WB

Unknown

Western blot - Anti-DKK1 antibody [EPR4759] (AB109416)

Blocking/Diluting buffer and concentration 5% NFDM /TBST

Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-DKK1 antibody [EPR4759] (ab109416) at 1/1000 dilution

All lanes:

His-Tagged human DKK1 (aa 32 to 266) recombinant protein, 100ng

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 1/20000 dilution

Predicted band size: 29 kDa

false