Anti-Wnt11 antibody [EPR25633-141] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Wnt11 antibody [EPR25633-141] - BSA and Azide free (AB316739)

This data was developed using ab316738, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized TF-1 (human erythroleukemia erythroblast) cells labelling Wnt11 with ab316738 at 1/50 (9.78 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing cytoplamic staining in TF-1 cell line. Negative control : A549.
Nuclear DNA was labeled with DAPI. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). 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 (2ug/ml) dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Wnt11 antibody [EPR25633-141] - BSA and Azide free (AB316739)

This data was developed using ab316738, the same antibody clone in a different buffer formulation.

Wnt11 was immunoprecipitated from 0.35 mg TF-1 (human Erythroleukemia erythroblast) whole cell lysate with ab316738 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316738 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : TF-1 (human Erythroleukemia erythroblast) whole cell lysate
Lane 2 : ab316738 IP in TF-1 (human Erythroleukemia erythroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316738 in TF-1 whole cell lysate

All lanes:

Immunoprecipitation - Anti-Wnt11 antibody [EPR25633-141] (<a href='/en-us/products/primary-antibodies/wnt11-antibody-epr25633-141-ab316738'>ab316738</a>) at 1/30 dilution

All lanes:

TF-1 (human Erythroleukemia erythroblast) 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

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-Wnt11 antibody [EPR25633-141] - BSA and Azide free (AB316739)

This data was developed using ab316738, the same antibody clone in a different buffer formulation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Wnt11 antibody [EPR25633-141] (<a href='/en-us/products/primary-antibodies/wnt11-antibody-epr25633-141-ab316738'>ab316738</a>) at 1/1000 dilution

Lane 1:

Untreated JEG-3 (human placenta epithelial cell) whole cell lysate at 60 µg

Lane 2:

JEG-3 treated with 300 ng/ml Brefeldin A for 24h whole cell lysate at 60 µg

Secondary

All lanes:

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

Observed band size: 39 kDa,124 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-Wnt11 antibody [EPR25633-141] - BSA and Azide free (AB316739)

This data was developed using ab316738, the same antibody clone in a different buffer formulation.

Negative control : NCI-H1975 (PMID : 31231131), A549 (PMID : 31231131).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Exposure time : Lane 1-2 : 3 seconds, Lane 3-4 : 26 seconds

All lanes:

Western blot - Anti-Wnt11 antibody [EPR25633-141] (<a href='/en-us/products/primary-antibodies/wnt11-antibody-epr25633-141-ab316738'>ab316738</a>) at 1/1000 dilution

Lane 1:

TF-1 (human Erythroleukemia erythroblast) whole cell lysate at 60 µg

Lane 2:

NCI-H1975 (human adenocarcinoma lung epithelial cell) whole cell lysate at 60 µg

Lane 3:

Saos-2 (human osteosarcoma epithelial cell) whole cell lysate at 60 µg

Lane 4:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 60 µg

Secondary

All lanes:

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

Observed band size: 39 kDa,36 kDa,124 kDa

false

• WB

Supplier Data

Western blot - Anti-Wnt11 antibody [EPR25633-141] - BSA and Azide free (AB316739)

This data was developed using ab316738, the same antibody clone in a different buffer formulation.

Low expression : lung (PMID : 15084607).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

Exposure time : Lane 1-2 : 37 seconds, Lane 3-4 : 26 seconds

All lanes:

Western blot - Anti-Wnt11 antibody [EPR25633-141] (<a href='/en-us/products/primary-antibodies/wnt11-antibody-epr25633-141-ab316738'>ab316738</a>) at 1/1000 dilution

Lane 1:

Rat fetal lung tissue lysate at 60 µg

Lane 2:

Rat lung tissue lysate at 60 µg

Lane 3:

Rat embryo tissue lysate at 60 µg

Lane 4:

Mouse embryo tissue lysate at 60 µg

Secondary

All lanes:

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

Observed band size: 39 kDa,15 kDa

false