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9966
IKK Isoform Antibody Sampler Kit

IKK Isoform Antibody Sampler Kit #9966

Western Blotting Image 1

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

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Western Blotting Image 2

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

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Western Blotting 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).

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Western Blotting Image 4

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

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Western Blotting 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.

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Flow Cytometry Image 6

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.

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Western Blotting 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).

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IF-IC Image 8

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).

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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 M R Mk 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 

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.

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.

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.

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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  13. Delhase, M. et al. (1999) Science 284, 309-13.
Entrez-Gene Id
1147 , 3551 , 9641 , 8517
Swiss-Prot Acc.
O15111 , O14920 , Q14164 , Q9Y6K9
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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