Anti-TIFA (phospho T9) antibody [EPR19853] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-TIFA (phospho T9) antibody [EPR19853] - BSA and Azide free (AB236848)

TIFA (phospho T9) was immunoprecipitated from 0.35 mg HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with ab214815 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214815 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours whole cell lysate 10 μg (Input).
Lane 2 : ab214815 IP in HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab214815 in HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours whole cell lysate (-).

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 3 minutes.

The lysates were kindly provided by our collaborator Dr Feng Shao's lab, NIBS.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214815).

All lanes:

Immunoprecipitation - Anti-TIFA (phospho T9) antibody [EPR19853] (<a href='/en-us/products/primary-antibodies/tifa-phospho-t9-antibody-epr19853-ab214815'>ab214815</a>)

Predicted band size: 21 kDa,43 kDa

Observed band size: 16 kDa,42 kDa,43 kDa

false

• WB

Collaborator

Western blot - Anti-TIFA (phospho T9) antibody [EPR19853] - BSA and Azide free (AB236848)

Blocking/Dilution buffer : 5% milk/TBST.

The image was kindly provided by our collaborator Dr Feng Shao's lab, NIBS.

S. flexneri infection induces an innate immune response (PMID : 28222186).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab214815).

All lanes:

Western blot - Anti-TIFA (phospho T9) antibody [EPR19853] (<a href='/en-us/products/primary-antibodies/tifa-phospho-t9-antibody-epr19853-ab214815'>ab214815</a>) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) transfected with 3×Flag-tagged TIFA expression vector for 28 hours, whole cell lysate at 20 µg

Lane 2:

HeLa transfected with 3×Flag-tagged TIFA T9A mutant expression vector for 28 hours, whole cell lysate at 20 µg

Lane 3:

HeLa transfected with 3×Flag-tagged TIFA expression vector for 24 hours, and then infected with S. flexneri 2457T for 4 hours, whole cell lysate at 20 µg

Lane 4:

HeLa transfected with 3×Flag-tagged TIFAT9A mutant expression vector for 24 hours, and then infected with S. flexneri 2457T for 4 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/5000 dilution

Predicted band size: 21 kDa

Observed band size: 26 kDa

false

Exposure time: 30s

• Dot

Supplier Data

Dot Blot - Anti-TIFA (phospho T9) antibody [EPR19853] - BSA and Azide free (AB236848)

Dot blot analysis of TIFA (phospho T9) labeled with ab214815 at 1/1000 dilution.

Lane 1 : TIFA (phospho T9) peptide (aa3-12).

Lane 2 : TIFA (phospho T9) peptide (aa5-15).

Lane 3 : TIFA non-phospho peptide (aa3-15).

Secondary used was Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214815).