IKK E (phospho-T501) antibody

• Catalog Number: orb345377
• Category: Antibodies
• Description: IKK E (phospho-T501) antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: IKKe phospho peptide corresponding to a region of the human protein surrounding pT501 conjugated to KLH.
• Concentration: 1 mg/mL
• Dilution range: ELISA: 1:5,000 - 1:25,000, WB: 1:500 - 1:3,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-IKKε pT501 antibody was affinity purified from monospecific antiserum by immunoaffinity purification against the phosphopeptide and cross adsorption against the non-phosphorylated form of the peptide followed by non-adsorption against a non-specific peptide backbone to further reduce non-specific reactivity. This phospho specific polyclonal antibody is specific for phosphorylated pT501 human IKKe. Reactivity with non-phosphorylated IKKe is minimal. Cross reactivity with pT501 phosphorylated IKKe from mouse, rat or other species has not been determined.
• Conjugation: Unconjugated
• UniProt ID: Q14164
• NCBI: Q14164.1
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.1% (w/v) Sodium Azide
• Alternative names: rabbit anti-IKK epsilon pT501 antibody, rabbit ant
• Note: For research use only
• Application notes: IKKε pT501 antibody is tested in ELISA, western blotting, and although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. An 85 kDa band corresponding to human IKKe is detected. HeLa cells or TNF inducible KBM-5 cells can be used as a positive control. Researchers should determine optimal titers for other applications.
• Expiration Date: 12 months from date of receipt.

IFI44 decreases the kinase activity of IKKβ and IKKε. Human 293T cells were silenced for IFI44, or for FKBP5, and were transfected with plasmids expressing His-IKKε (A) or MYC-IKKβ (B), together with IFI44-HA, and FKBP5-FLAG expression plasmids. At 24 hpt, IKKε (A) and IKKβ (B) complexes were purified with anti-His and anti-MYC antibodies, respectively, and these complexes were assayed in kinase assays using IRF-3 (for the IKKε complexes shown in panel A) and IkBα (for the IKKβ complexes shown in panel B) as substrates. The levels of phosphorylated and unphosphorylated forms of IRF-3 (panel A, bottom blot) and IkBα (panel B, third and fourth blots) were analyzed by Western blotting using specific antibodies. Levels of IKKε were analyzed using an anti-His-specific antibody (A, first blot) and anti-pIKKε (A, second blot), and levels of IKKβ were analyzed using an anti-MYC-specific antibody (B, first blot) and anti-pIKKβ (B, second blot). Protein expression levels in cells expressing IKKε (A) and IKKβ (B) alone were assigned a value of 100% for comparisons with the levels of expression in cells expressing the different combinations of IKKε/IFI44/FKBP5 (A) or IKKβ/IFI44/FKBP5 (B) (numbers are indicated below each plot). pIRF-3 and IRF-3 levels (observed in the same bottom blot in panel A) and pIkBα and IkBα (third and bottom blot in panel B) are represented with numbers below each blot. Levels of pIRF-3 and pIkBα normalized to the levels of IKKε and IKKβ are represented in the bottom graphs in panels A and B, respectively. Molecular weight markers are indicated (in kilodaltons) on the right.