Anti-TZP antibody [EPR25998-35] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-TZP antibody [EPR25998-35] - BSA and Azide free (AB285369)

This data was developed using ab285366, the same antibody clone in a different buffer formulation.

TZP was immunoprecipitated from 0.35 mg HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate 10 μg with ab285366 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab285366 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HCT116 whole cell lysate 10 μg

Lane 2 : ab285366 IP in HCT116 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab285366 in HCT116 whole cell lysate

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

Exposure time : 3.25 seconds.

All lanes:

Immunoprecipitation - Anti-TZP antibody [EPR25998-35] (<a href='/en-us/products/primary-antibodies/tzp-antibody-epr25998-35-ab285366'>ab285366</a>)

Predicted band size: 115 kDa

false

• WB

Lab

Western blot - Anti-TZP antibody [EPR25998-35] - BSA and Azide free (AB285369)

This data was developed using ab285366, the same antibody clone in a different buffer formulation.

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

Negative control : NCI-H460 (PMID : 26722404)

Exposure time : 8 seconds

All lanes:

Western blot - Anti-TZP antibody [EPR25998-35] (<a href='/en-us/products/primary-antibodies/tzp-antibody-epr25998-35-ab285366'>ab285366</a>) at 1/1000 dilution

Lane 1:

HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

NCI-H460 (human Large Cell Lung Cancer 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 1/100000 dilution

Predicted band size: 115 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-TZP antibody [EPR25998-35] - BSA and Azide free (AB285369)

This data was developed using ab285366, the same antibody clone in a different buffer formulation.

Lane 2 : This blot was developed using a higher sensitivity ECL substrate.

Exposure time : Lane 1 : 15 seconds; Lane 2 : 3 minutes.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-TZP antibody [EPR25998-35] (<a href='/en-us/products/primary-antibodies/tzp-antibody-epr25998-35-ab285366'>ab285366</a>) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Mouse heart tissue 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

Predicted band size: 115 kDa

Observed band size: 140 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TZP antibody [EPR25998-35] - BSA and Azide free (AB285369)

This data was developed using ab285366, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 cells labelling TZP with ab285366 at 1/1000 (0.589 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 μg/ml) dilution (Green).

Confocal image showing nuclear staining in HCT 116 cell line.

Negative control : NCI-H460 (PMID : 26722404).

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

• IP

Supplier Data

Immunoprecipitation - Anti-TZP antibody [EPR25998-35] - BSA and Azide free (AB285369)

This data was developed using ab285366, the same antibody clone in a different buffer formulation.

TZP was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate 10 μg with ab285366 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab285366 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : F9 whole cell lysate 10 μg

Lane 2 : ab285366 IP in F9 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab285366 in F9 whole cell lysate

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

Exposure time : 3.25 seconds.

All lanes:

Immunoprecipitation - Anti-TZP antibody [EPR25998-35] (<a href='/en-us/products/primary-antibodies/tzp-antibody-epr25998-35-ab285366'>ab285366</a>)

Predicted band size: 115 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TZP antibody [EPR25998-35] - BSA and Azide free (AB285369)

This data was developed using ab285366, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized F9 cells labelling TZP with ab285366 at 1/1000 (0.589 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 μg/ml) dilution (Green).

Confocal image showing nuclear staining in F9 cell line is observed.

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