Intended Use:The Phospho-IKKa/b (Ser176/180) Antibody Sampler Kit contains reagents to examine protein levels of IKKa when phosphorylated at Ser176/180 and IKKb when phosphorylated at Ser177/181. The kit contains primary and secondary antibodies to perform four Western Blots with each antibody.
Background:The NF-κB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins (1-3). Most agents that activate NF-κB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degrada- tion of IκB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IκB kinase (IKK) complex, whose catalysis is generally carried out by three tightly associated IKK subunits. IKKa and IKKb serve as the catalytic subunits of the kinase. IKKγ serves as the regulatory subunit (8-9). Activation of IKK depends on phosphorylation; Ser177 and Ser181 in the activation loop of IKKb (176 and 180 in IKKa) are the specific sites whose phosphorylation causes conformational changes resulting in kinase activation (10-13).
Kit Components:1. Phospho-IKKa (Ser176)/IKKb (Ser177) (C84E11) Mab rabbit x human, 1x20ul2. Phospho-IKKa/b (Ser176/180) (16A6) Mab rabbit x human, 1x20ul3. Phospho-IKKa/b (Ser176/180) Pab rabbit x human, 1x20ul4. IgG (HRP) Pab Goat x Rabbit, 1x100ul
Applications:Suitable for use in Western Blot. Phospho-IKKa/b (Ser176/180) (16A6) Mab rabbit x human has been tested in Immunohistochemistry (frozen and paraffin). Other applications not tested.
Recommended Dilution:Western Blot: 1:1000Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size:1Kit
Grade:Affinity Purified
Purity:Purified by protein A and peptide affinity chromatography.
Form:Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.
Specificity:Recognizes IKKa at 85kD only when phosphorylated at Ser176/180 and IKKb at 87kD only when phosphorylated at Ser177/181.1. Phospho-IKKa (Ser176)/IKKb (Ser177) Mab rabbit x human: recognizes human, mouse. Sequence homology: rat, monkey, bovine.2. Phospho-IKKa/b (Ser176/180) Mab rabbit x human: recognizes human, mouse, rat, monkey. Sequence homlogy: bovine.3. Phospho-IKKa/b (Ser176/180) Pab rabbit x human recognzies human, mouse, rat, monkey
Calc Applications Abbrev:WB
Calc Crossreactivity:Hu Mk Mo Rt
Immunogen:Synthetic phosphopeptide corresponding to residues surrounding Ser176/180 of human IKKa and Ser177/181 of IKKb, respectively, and are purified by protein A and peptide affinity chromatography.