Antibodies, Proteins, Kits and Reagents for Life Science

288 134 ₸ Product size

100 µl 288 134 ₸ 10 µl 117 264 ₸

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Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19853] to TIFA (phospho T9)
Suitable for: WB, Dot blot, IP
Reacts with: Human

Product name

Anti-TIFA (phospho T9) antibody [EPR19853]
See all TIFA primary antibodies
Description

Rabbit monoclonal [EPR19853] to TIFA (phospho T9)
Host species

Rabbit
Tested Applications & Species

Application Species
IP
Human

WB
Human

Recombinant fragment

See all applications and species data
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: 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. 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. IP: 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.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Application	Species
IP	Human
WB	Human Recombinant fragment

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19853
Isotype

IgG
Research areas

Western blot - Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815)

All lanes : Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815) 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
Lane 2 : HeLa transfected with 3×Flag-tagged TIFA T9A mutant expression vector for 28 hours, whole cell lysate
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
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

Lysates/proteins at 20 µg per lane.

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
why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% milk/TBST.

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

S. flexneri infection induces an innate immune response (PMID: 28222186).
Immunoprecipitation - Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815)

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 1/5,000 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 cell lysates were kindly provided by our collaborator Dr. Feng Shao, NIBS.
Western blot - Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815)

All lanes : Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815) at 1/5000 dilution

Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-tagged TIFA expression vector, whole cell lysate
Lane 2 : 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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 21 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?

Exposure time: 103 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The band observed represents GFP-tagged TIFA pT9.

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

B. cenocepacia J2315 infection elicits an inflammatory response. (PMID: 27133449)
Dot Blot - Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815)

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.
Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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