Anti-DKK1 antibody [EPR26160-102]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DKK1 antibody [EPR26160-102] (AB307367)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling DKK1 with ab307367 at 1/50 (10.78 ug/ml) dilution, followed by ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line. Low expression : C2C12 (PMID : 18305214).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-DKK1 antibody [EPR26160-102] (AB307367)

DKK1 was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate 10 ug with ab307367 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307367 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate 10 ug
Lane 2 : ab307367 IP in Neuro-2a whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307367 in Neuro-2a whole cell lysate

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

Exposure time : 3 seconds

The band beneath the target band is likely to be degraded target fragments.

All lanes:

Immunoprecipitation - Anti-DKK1 antibody [EPR26160-102] (ab307367) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate 10 μg

Lane 2:

Neuro-2a whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab307367 in Neuro-2a whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 35 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-DKK1 antibody [EPR26160-102] (AB307367)

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

Low expression : C2C12 (PMID : 18305214), 3T3-L1 (PMID : 19943155), HEK-293 (PMID : 18502762), Jurkat (PMID : 20653953)

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The identity of the higher MW band at approximately 75 kDa is unknown.

The molecular weight observed is consistent with what has been described in the literature (PMID : 14695885).

Exposure time :

Lane 1-3 : 37 seconds

Lane 4-6 : 81 seconds

All lanes:

Western blot - Anti-DKK1 antibody [EPR26160-102] (ab307367) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 3:

3T3-L1 (mouse embryonic fibroblast) differentiated into adipocyte-like cells whole cell lysate at 20 µg

Lane 4:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysat at 20 µg

Lane 5:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 20 μg at 20 µg

Lane 6:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) 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

Observed band size: 35 kDa

true

• WB

Supplier Data

Western blot - Anti-DKK1 antibody [EPR26160-102] (AB307367)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTLysates were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the higher MW band at approximately 75 kDa is unknown. The molecular weight observed is consistent with what has been described in the literature (PMID : 14695885). Exposure time : Lane 1-3 : 37 seconds, Lane 4-6 : 81 seconds.

All lanes:

Western blot - Anti-DKK1 antibody [EPR26160-102] (ab307367) at 1/1000 dilution

All lanes:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 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

Observed band size: 35 kDa

false

Exposure time: 158s