Revision 1

#64662

Store at -20C

NF-κB Pathway Antibody Sampler Kit II

1 Kit

(9 x 20 microliters)

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Orders:

877-616-CELL (2355)

orders@cellsignal.com

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
IKKα (D3W6N) Rabbit mAb6129420 µl85 kDaRabbit IgG
IKKβ (D30C6) Rabbit mAb894320 µl87 kDaRabbit IgG
Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb269720 µl85 IKK-alpha 87 IKK-beta kDaRabbit IgG
IκBα (44D4) Rabbit mAb481220 µl39 kDaRabbit IgG
Phospho-IκBα (Ser32) (14D4) Rabbit mAb285920 µl40 kDaRabbit IgG
NF-κB p65 (D14E12) XP® Rabbit mAb824220 µl65 kDaRabbit IgG
Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb303320 µl65 kDaRabbit IgG
NF-κB1 p105/p50 (D4P4D) Rabbit mAb1358620 µl50 Active form. 120 Precursor kDaRabbit IgG
Phospho-NF-κB p65 (Ser529) Antibody9687420 µl65 kDaRabbit
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The NF-κB Pathway Antibody Sampler Kit II contains reagents to examine the activation state and total protein levels of key proteins in the NF-κB pathway: IKKα, IKKβ, NF-κB p65/RelA, and IκBα. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

The transcriptional nuclear factor κB (NF-κB)/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins. Activation occurs via phosphorylation of IκBα at Ser32 and Ser36, resulting in the ubiquitin-mediated proteasome-dependent degradation of IκBα and the release and nuclear translocation of active NF-κB dimers. The regulation of IκBβ and IκBε is similar to that of IκBα, however, the phosphorylation and degradation of these proteins occurs with much slower kinetics. Phosphorylation of IκBβ occurs at Ser/Thr19 and Ser23, while IκBε can be phosphorylated at Ser18 and Ser22. The key regulatory step in this pathway involves activation of a high molecular weight IkappaB kinase (IKK) complex, consisting of three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase. Activation of IKK depends on phosphorylation at Ser177 and Ser181 in the activation loop of IKKβ (176 and 180 in IKKα). NF-κB-inducing kinase (NIK), TANK-binding kinase 1 (TBK1), and its homolog IKKε (IKKi), phosphorylate and activate IKKα and IKKβ.
The NF-κB family of transcription factors is comprised of five proteins in mammals, p65/RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). p105 and p100 are proteolytically processed to produce p50 and p52, respectively. The 50 kDa active form is produced through proteolytic processing following IKK-mediated phosphorylation of p105 at multiple sites (Ser922, 924, 928, and 933), while p100's processing to p52 is induced by phosphorylation of Ser864 and Ser868. The p50 and p52 products form dimeric complexes with Rel proteins, which are then able to bind DNA and regulate transcription. Phosphorylation of p65/RelA at Ser276 by PKA C and MSK1 enhances transcriptional activity. p65 phosphorylation at Ser536 regulates activation, nuclear localization, protein-protein interactions, and transcriptional activity. PMA-induced NF-κB transcriptional activity is dependent on the region of p65 containing the potential phosphorylation sites Ser457, Thr458, Thr464, and Ser468. Phosphorylation of Ser468 by GSK-3β inhibits basal p65 activity.

Background References

Trademarks and Patents

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Limited Uses

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Orders: 877-616-CELL (2355) • orders@cellsignal.com • Support: 877-678-TECH (8324) • info@cellsignal.com • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.