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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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.

Product Citations
1. p38-dependent marking of inflammatory genes for increased NF-kB recruitment. Saccani S, S Pantano, G Natoli. Nature Immunology 3:69-75 (2002). 
WB (human purified monocytes):  Figs. 4c (bottom panel), 5a (bottom panel), 5c.
2. Two waves of nuclear factor kB recruitment to target promoters. Saccani S, S Pantano, G Natoli. J. Exp. Med. 193:1351-1359 (2001).
WB (mouse LPS-stimulated Raw 264.7 macrophages), Fig. 1.
3. Modulation of NF-kB activity by exchange of dimers. Saccani S, S Pantano, G Natoli. Molecular Cell 11:1563-1574 (2003).
WB (HEK-293 cells transfected with different NF-kB subunits) Fig. 1b
WB (human dendritic cells) Fig. 2a
WB (mutant IkBa adenovirus infected dendritic cells) Fig. 2c
4. PC-SPES: A potent inhibitor of nuclear factor-κB rescues mice from lipopolysaccharide-induced septic shock. Ikezoe T, Y Yang, D Heber, H Taguchi, H P Koeffler. Molecular Pharmacology 64:1521-1529 (2003).  Imgenex antibodies cited [WB, LPS-treated RAW 264.7 cells]:
1.  IkBa (Ser 32/Ser 36) [IMG-156A], Fig. 3a

2.  IkBa [IMG-127A], Fig. 3b
5. Degradation of promoter-bound p65/RelA is essential for the prompt termination of the nuclear factor kB response. Saccani S, I Marazzi, A A Beg, G Natoli. J. Exp. Med., 200:107-113 (2004).
WB (TNF-a stimulated WT and IkBa-/- NIH 3T3), Fig. 1a
     Note: IkBa (IMG-127A) recognized the IkBa wildtype but not the IkBa-/-, as expected
WB (p65 transfected HEK-293 cells), Fig. 2a
WB (NIH 3T3), Figs. 4a, 4d
IP (p65 transfected 293T cells), Fig. 2b
     Note: IkBa (IMG-127A) co-IP'd p65
6. Andrographolide attenuates inflammation by inhibition of NF-kB activation through covalent modification of reduced cysteine 62 of p50. Xia Y F, B Q Ye, Y D Li, J G Wang, X J He, X Lin, X Yao, D Ma, A Slungaard, R P Hebbel, N S. Key, J Geng. Journal of Immunology 173:4207-4217 (2004).
WB (TNF-a treated 293 cells), Fig 6a
7. Differential effects of NF-kB on apoptosis induced by DNA-damaging agents: the type of DNA damage determines the final outcome. Strozyk E, B Pöppelmann, T Schwarz, D Kulms. Oncogene 25:6239-6251 (2006).  
WB (human oral epithelial carcinoma cell line KB) Figs. 1a, 5b, 5d
8. NF-kB Is required for apoptosis prevention during herpes simplex virus type 1 infection. Goodkin M L, A T Ting, J A Blaho. Journal of Virology, 77:7261-7280 (2003).
WB (mouse HEp-2 cells and clonal CD-14 transfected cells treated with either TNF-a or TNF-a plus CHX), Fig. 9a
WB (mouse HEp-2 cells treated with TNF-a plus CHX and mutant IkBaDN cells treated with TNF-a), Fig. 9b
WB (mouse mutant HEp-2 and IkBaDN cells infected with HSV-1(F)), Fig. 10b
WB (mouse mutant HEp-2 and IkBaDN cells infected with HSV-1(KOS1.1)), Fig. 10d
9. The IkB kinase is a key factor in triggering influenza A virus-induced inflammatory cytokines production in airway epithelial cells. Bernasconi D, C Amici, S L Frazia, A Ianaro, M G Santoro. Journal of Biological Chemistry 280:24127-24134 (2005).
WB (WSN-infected A549 human pulmonary cells), Fig. 1a bottom
10. Selected Toll-like receptor agonist combinations synergistically trigger a T helper type 1−polarizing program in dendritic cells. Napolitani G, A Rinaldi, F Bertoni, F Sallusto, A Lanzavecchia. Nature Immunology  6:769-776 (2005).
WB (human MoDC stimulated with LPS, R848, and poly(I:C)), Fig. 8b
11. B Cell maturation antigen, the receptor for a proliferation-inducing ligand and B Cell-activating factor of the TNF family, induces antigen presentation in B Cells. Yang M, H Hase, D Legarda-Addison, L Varughese, B Seed, A T. Ting. Journal of Immunology, 175:2814-2824 (2005).Imgenex antibodies cited [WB]:
1. IkBa Ser 32/Ser 36 [IMG-156A], (A20 B cells stimulated with APRIL), Fig. 2c

2. IkBa [IMG-127A], (A20 cells stimulated with anti-TNFR2), Fig. 6a
12. A cyanobacterial LPS antagonist prevents endotoxin shock and blocks sustained TLR4 stimulation required for cytokine expression Macagno A, M Molteni, A Rinaldi, F Bertoni, A Lanzavecchia, C Rossetti, F Sallusto. J. Exp. Med., 203:1481-1492 (2006).
WB (DCs stimulated with LPS and treated with CyP), Figs. 4a, 6e
13. NF-kB and COX-2 during oral tumorigenesis and in assessment of minimal residual disease in surgical margins. Santhi W S, P Sebastian, B T Varghese, O Prakash, M R Pillai. Experimental and Molecular Pathology 81:123-130 (2006).  Imgenex products used (human):
1.  IkBa [IMG-127A]: IHC paraffin (normal oral mucosa, leukoplakia, and oral cancer), Fig. 1
IHC paraffin analysis (normal oral mucosa, leukoplakia, and oral cancer), Fig. 2
IHC paraffin (oral cancer: primary tumor, surgical margin, and extra margin tissue), Fig 6
IHC paraffin analysis (oral cancer: primary tumor, surgical margin, and extra margin tissue), Fig. 7

2.  p65 [IMG-150A]: IHC paraffin (normal oral mucosa, leukoplakia, and oral cancer), Fig. 1
IHC paraffin analysis (normal oral mucosa, leukoplakia, and oral cancer), Fig. 2
IHC paraffin (oral cancer: primary tumor, surgical margin, and extra margin tissue), Fig. 6
IHC paraffin analysis (oral cancer: primary tumor, surgical margin, and extra margin tissue), Fig. 7

3.  p50 [IMG-5812A]: IHC paraffin (normal oral mucosa, leukoplakia, and oral cancer), Fig. 1
IHC paraffin analysis (normal oral mucosa, leukoplakia, and orla cancer), Fig. 2
IHC paraffin (oral cancer: primary tumor, surgical margin, and extra margin tissue), Fig. 6
IHC paraffin amalysis (oral cancer: primary tumor, surgical margin, and extra margin tissue), Fig. 7

4. NF-kB ELISA [IMK-503]: (normal oral mucosa, leukoplakia, and oral cancer), Fig. 3
(oral cancer: primary tumor, surgical margin, and extra margin tissue), Fig. 8
14. Paeonol attenuates TNBS-induced colitis by inhibiting NF-κB and STAT1 transactivation. Ishiguro K, T Ando, O Maeda, M Hasegawa, K Kadomatsu, N Ohmiya, Y Niwa, R Xavier, H Goto. Toxicology and Applied Pharmacology, 217:35-42 (2006).
WB (mouse colon tissue CW-2 cells stimulated with TNFa in presence of Paeonol)  Fig. 5a
15. BAFF and APRIL support chronic lymphocytic leukemia B cell survival through activation of the canonical NF-kB pathway.  Endo T, M Nishio, T Enzler, H B Cottam, T Fukuda, D F James, M Karin, T J Kipps. Blood 109:703-710 (2007).
WB (human CLL B cells cultured with rhBAFF or rhAPRIL), Fig. 2b
WB (CLL cells transfected with HA-tagged IkBaSR or empty pcDNA3 vector stimulated with rhBAFF), Fig. 5a
16. Targeting the NF-κB signaling pathway in Notch1-induced T-cell leukemia.  T Vilimas, J Mascarenhas, T Palomero, M Mandal, S Buonamici, F Meng, B Thompson, C Spaulding, S Macaroun, M L Alegre, B L Kee, A Ferrando, L Miele, I Aifantis. Nature Medicine 13, 70-77 (2007).
IP (CEM cells treated with Z-Leu-Leu-Nle-CHO and LY411575), Fig. 2d
WB (CEM cells IP'd with IKK antibody or Notch1-IC antibody), Figs. 2e, 2f
17. DHMEQ, a novel nuclear factor-κB inhibitor, induces selective depletion of alloreactive or phytohaemagglutinin-stimulated peripheral blood mononuclear cells, decreases production of T helper type 1 cytokines, and blocks maturation of dentritic cells.  Nishioka C, T Ikezoe, Y Jing, K Umezawa, A Yokoyama.  Immunology 124:198-205 (2008).
WB (acute lymphoblastic T-cell leukemia Jurkat cells pretreated with DHMEQ and exposed to phytohaemagglutin), Fig. 4.
18.