Anti-IRX5 antibody [EPR22724-43] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-IRX5 antibody [EPR22724-43] - BSA and Azide free (AB288301)

This data was developed using ab246494, the same antibody clone in a different buffer formulation.

IRX5 was immunoprecipitated from 0.35 mg 293T transfected with a DDDDK-tagged mouse IRX5 (WT) expression vector, whole cell lysate 4 ug with ab246494 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246494 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 293T transfected with a DDDDK-tagged mouse IRX5 (WT) expression vector, whole cell lysate 4 ug

Lane 2 : 293T transfected with a DDDDK-tagged mouse IRX5 (WT) expression vector, whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246494 in 293T transfected with a DDDDK-tagged mouse IRX5 (WT) expression vector, whole cell lysate

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

Exposure time : 1 second

All lanes:

Immunoprecipitation - Anti-IRX5 antibody [EPR22724-43] (<a href='/en-us/products/primary-antibodies/irx5-antibody-epr22724-43-ab246494'>ab246494</a>)

Predicted band size: 50 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-IRX5 antibody [EPR22724-43] - BSA and Azide free (AB288301)

This data was developed using ab246494, the same antibody clone in a different buffer formulation.

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

In lane 1, the lysate was stored at -80°C prior to Western Blotting. In lane 2, lysate was freshly made and used for Western Blotting immediately to minimize protein degradation.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-IRX5 antibody [EPR22724-43] (<a href='/en-us/products/primary-antibodies/irx5-antibody-epr22724-43-ab246494'>ab246494</a>) at 1/1000 dilution

Lane 1:

Mouse lung (frozen) tissue lysate at 20 µg

Lane 2:

Mouse lung (fresh) tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-IRX5 antibody [EPR22724-43] - BSA and Azide free (AB288301)

This data was developed using ab246494, the same antibody clone in a different buffer formulation.

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

Exposure time : 3 seconds

All lanes:

Western blot - Anti-IRX5 antibody [EPR22724-43] (<a href='/en-us/products/primary-antibodies/irx5-antibody-epr22724-43-ab246494'>ab246494</a>) at 1/1000 dilution

Lane 1:

HeLa transfected with a DDDDK-tagged mouse IRX3 (WT) expression vector, whole cell lysate at 4 µg

Lane 2:

HeLa transfected with a DDDDK-tagged mouse IRX5 (WT) expression vector, whole cell lysate at 4 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 50 kDa

Observed band size: 60 kDa

false

• WB

Lab

Western blot - Anti-IRX5 antibody [EPR22724-43] - BSA and Azide free (AB288301)

This data was developed using ab246494, the same antibody clone in a different buffer formulation.

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

Exposure time : 15 seconds

The image is kindly provided by an anonymous collaborator.

All lanes:

Western blot - Anti-IRX5 antibody [EPR22724-43] (<a href='/en-us/products/primary-antibodies/irx5-antibody-epr22724-43-ab246494'>ab246494</a>) at 1/1000 dilution

Lane 1:

IRX5 (WT) over expression whole cell lysate

Lane 2:

Mouse lung (WT) tissue lysate

Lane 3:

IRX5 knock out mouse lung tissue lysate

Predicted band size: 50 kDa

Observed band size: 60 kDa

false