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Monoclonal Antibody to IkBa (Clone 6A920) |
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| Catalog No: |
IMG-127A |
| Contents: |
100ug in 200ul PBS containing 0.05% BSA, 0.05% sodium azide. Sodium azide is highly toxic. |
| Isotype: |
Mouse IgG1, Kappa |
| Clone: |
6A920 |
| Purification: |
Protein G Chromatography |
| Species: |
Human, Mouse |
| Host: |
Mouse |
| Application: |
Flow (Intracellular): 0.25-1 ug/10^6 cells
IHC (paraffin): see Santhi et. al. 2006 for details
IP: 1 ug/ml
Western blot analysis: 1-2 ug/ml |
| Storage: |
Store at 4°C. For long-term storage, store at -20°C. |
Recommended
Positive Control: |
Daudi, Jurkat, or NIH 3T3 |
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Background
NF-kB is silenced in the cytoplasm by an inhibitory protein, IkB (1). Synthesis of IkBa is autoregulated (2). IkB proteins are phosphorylated by IkB kinase complex consisting of at least three proteins, IKK1/a, IKK2/b, and IKK3/g (3-6). External stimuli such as tumor necrosis factor or other cytokines results in phosphorylation and degradation of IkB releasing NF-kB dimers. NF-kB dimer subsequently translocates to the nucleus and activates target genes. Six members of IkB family members have been identified (1). One of the first gene induced following NF-kB activation is IkBa. |
Antigen
A recombinant protein corresponding to amino acid residues 32-291 of human IkBa was used as immunogen. |
Application Notes
A 40 kDa band is observed. The antibody reacts with human and mouse IkBa. It may react with other species, but this has not been tested at IMGENEX. |
Genebank Info (Protein)
NP_065390 |
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Jurkat cells were treated for 30 min with 100 ug/ml ALLN (N-Acetyl-Leu-Leu-Norleucinal; a Calpain inhibitor and also proteasome inhibitor that prevents IkBa dephosphorylation) followed by incubation with (lanes 2 & 4) or without 1 nM TNF-a (1 & 3). The membranes were blotted with IMG-156 (lanes 1 & 2) or IMG-127 (that recognizes both non-phospho and phosphorylated forms of IkBa) and immunoreactivity was detected by ECL. The data shows that IMG-156 detects specifically the phosphorylated form of IkBa. |
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Western blot analysis of IkBa using IMG-127 at 2 ug/ml in (A) Daudi and (B) NIH 3T3 whole cell lysate. |
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Intracellular staining of 10^6 ThP-1 cells using 0.25 ug of IMG-127A. Shaded histogram represents cells alone, green represents the isotype control (IMGENEX, 20109), and red represents the IkBa antibody. IMGENEX's intracellular flow kit (IMGENEX, 10083K) was used for this test, and an anti-mouse IgG FITC conjugated secondary. |
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Reference
1. Verma, I.M., Genes. Dev. 9: 2723-2735, 1995.
2. Verma, I. And Stevenson, J.K. Proc. Nat. Acad. Sci USA 94: 11758 (1997).
3. DiDonato, J.A., et al. Nature 388: 548- (1997).
4. Regnier, C.H., et al. Cell 90, 373 (1997).
5. Mercurio, F., et al. Science 278: 860 (1997).
6. Yamaoka, S., et al. Cell 93: 1231-1240 (1998).
7. Sanjo H, Kawai T, and Akira S. DRAKs, novel serine/threonine kinases related to death associated protein kinase that trigger apoptosis. J Biol Chem 273 (44): 29066-29071, 1998.
8. Matsumoto M, Takeda K, Sanjo H, and Akira S. ZIP kinase, a novel serine/threonine kinase which mediates apoptosis. Mol Cell Biol 18 (3): 1642-1651, 1998.
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Research purposes only.
Not for diagnostic or in vivo use. This product is guaranteed to perform as indicated on the datasheet for one year from the date of purchase.
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