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9966
IKK Isoform Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

IKK Isoform Antibody Sampler Kit #9966

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IKK Isoform Antibody Sampler Kit: Image 1

Flow cytometric analysis of HCT 116 cells using IKKα (3G12) Mouse mAb (blue) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (red). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.

IKK Isoform Antibody Sampler Kit: Image 2

Western blot analysis of extracts from various cell lines using IKKβ (D30C6) Rabbit mAb.

IKK Isoform Antibody Sampler Kit: Image 3

Western blot analysis of extracts from HeLa cells, untreated or treated with TNF-α (20 ng/ml) and Calyculin A #9902 (50 nM), using Phospho-IKKγ (Ser376) Antibody #2689 (top) or IKKγ (DA10-12) Mouse mAb (bottom).

IKK Isoform Antibody Sampler Kit: Image 4

Western blot analysis of recombinant IKKε Kinase #7553 and RAW 264.7 cells, untreated or LPS-treated, using IKKε Antibody.

IKK Isoform Antibody Sampler Kit: Image 5

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

IKK Isoform Antibody Sampler Kit: Image 6

Confocal immunofluorescent analysis of HCT 116 (high expression; left) and IGROV-1 (low expression; right) cells using IKKα (3G12) Mouse mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

IKK Isoform Antibody Sampler Kit: Image 7

Western blot analysis of extracts from wild-type, IKKα (-/-), and IKKβ (-/-) mouse embryonic fibroblasts (MEFs) using IKKβ (D30C6) Rabbit mAb (upper) and GAPDH (14C10) Rabbit mAb #2118 (lower).

IKK Isoform Antibody Sampler Kit: Image 8

Western blot analysis of extracts from various cell lines using IKKα (3G12) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

IKK Isoform Antibody Sampler Kit: Image 9

Western blot analysis of HeLa cell extracts (lane 1) or IKKα knock-out HeLa cells (lane 2), using IKKα (3G12) Mouse mAb #11930 (upper), or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the knockout HeLa cells confirms specificity of the antibody for IKKα.

To Purchase # 9966T
Product # Size Price
9966T
1 Kit  (4 x 20 µl) $ 327

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
IKKα (3G12) Mouse mAb 11930 20 µl
  • WB
  • IF
  • F
H Mk 85 Mouse IgG1
IKKβ (D30C6) Rabbit mAb 8943 20 µl
  • WB
  • IP
H Mk M R 87 Rabbit IgG
IKKγ (DA10-12) Mouse mAb 2695 20 µl
  • WB
H R 50 Mouse IgG1
IKKε Antibody 2690 20 µl
  • WB
H M R 80 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

Product Description

The IKK Isoform Antibody Sampler Kit provides an economical means to investigate NFkappaB signaling within the cell. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.

Specificity / Sensitivity

IKKα, IKKβ, IKKγ, and IKKε antibodies detect endogenous levels of total IKKα, IKKβ, IKKγ and IKKε proteins, respectively. These antibodies do not cross-react with IKK subunits other than their specified target.

Source / Purification

The IKKβ (D30C6) Rabbit mAb was produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human IKKβ protein. The IKKα (3G12) Mouse mAb was produced by immunizing animals with a recombinant protein specific to a fragment of human IKKα protein. The IKKγ (DA10-12) Mouse mAb was produced by immunizing animals with full length human GST-IKKγ protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues at the carboxyl terminus of human IKKγ. Antibodies are purified by protein A and peptide affinity chromatography.

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 degradation 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. IKKα and IKKβ serve as the catalytic subunits of the kinase and IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends upon phosphorylation at Ser177 and Ser181 in the activation loop of IKKβ (Ser176 and Ser180 in IKKα), which causes conformational changes, resulting in kinase activation (10-13).

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  3. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
  4. Brown, K. et al. (1995) Science 267, 1485-8.
  5. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  8. Zandi, E. et al. (1997) Cell 91, 243-52.
  9. Karin, M. (1999) Oncogene 18, 6867-74.
  10. DiDonato, J.A. et al. (1997) Nature 388, 548-54.
  11. Mercurio, F. et al. (1997) Science 278, 860-6.
  12. Johnson, L.N. et al. (1996) Cell 85, 149-58.
  13. Delhase, M. et al. (1999) Science 284, 309-13.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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