Anti-Dkk3 antibody [EPR15611] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Dkk3 antibody [EPR15611] - BSA and Azide free (AB232657)

Clone EPR15611 (ab232657) has been successfully conjugated by Abcam. This image was generated using Anti-Dkk3 antibody [EPR15611] (Alexa Fluor® 488). Please refer to ab201262 for protocol details.

ab201262 staining Dkk3 in HepG2 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab201262 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IP

Supplier Data

Immunoprecipitation - Anti-Dkk3 antibody [EPR15611] - BSA and Azide free (AB232657)

Western blot analysis of Dkk3 in Human fetal brain lysate immunoprecipitated with ab186409 at 1/50 dilution (Lane 1). Lane 2 : PBS instead of Human fetal brain lysate.

Secondary antibody : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab186409).

All lanes:

Immunoprecipitation - Anti-Dkk3 antibody [EPR15611] (<a href='/en-us/products/primary-antibodies/dkk3-antibody-epr15611-ab186409'>ab186409</a>)

Predicted band size: 38 kDa,56 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-Dkk3 antibody [EPR15611] - BSA and Azide free (AB232657)

Blocking buffer and concentration : 5% NFDM/TBST  Diluting buffer and concentration : 5% NFDM/TBST DKK3 shows 50-70 kDa bands, which is due to glycosylation (PMID : 28674110. PMID : 28738084). DKK3 is not expressed in PC-3 and BxPC-3. Negative control : PC-3 (PMID : 22941469), BxPC-3 (PMID : 28470144, PMID : 26395974, PMID : 34391478). This data was developed using the same antibody clone in a different buffer formulation (ab186409).

All lanes:

Western blot - Anti-Dkk3 antibody [EPR15611] (<a href='/en-us/products/primary-antibodies/dkk3-antibody-epr15611-ab186409'>ab186409</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 38 kDa

false

Exposure time: 10s