Product Pathways - NF-kappaB Signaling
Phospho-NF-KappaB p65 (Ser536) (93H1) Rabbit mAb (Alexa Fluor® 647 Conjugate) #4887
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| F | H M (R) | Endogenous | Rabbit IgG |
Applications Key:
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-NF-KappaB p65 (Ser536) (93H1) Rabbit mAb (Alexa Fluor® 647 Conjugate) detects NF-kappaB p65 only when phosphorylated at serine 536. It does not cross-react with the p50 subunit or other related proteins.
Source / Purification
Monoclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser536 of human NF-kappaB p65. The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.
Flow Cytometry
Flow cytometric analysis of THP-1 cells, untreated (blue) or TNF-α treated (green), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Alexa Fluor® 647 Conjugate).
Description
Cell Signaling Technology Antibody conjugated to Alexa Fluor ® 647 fluorescent dye and tested in-house for direct Flow Cytometric analysis of human cells. The unconjugated antibody, #3033, reacts with Phospho-NF-KappaB p65 (Ser536) from human, mouse. and rat. CST expects that Phospho-NF-KappaB p65 (Ser536) (93H1) Rabbit mAb (Alexa Fluor® 647 Conjugate) will also recognize Phospho-NF-KappaB p65 (Ser536) in these species.
Background
Transcription factors of the nuclear factor kappa B (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50) and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through an ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which is then translocated to the nucleus (9-11).
- Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
- Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
- Haskill, S. et al. (1991) Cell 65, 1281-9.
- Thompson, J.E. et al. (1995) Cell 80, 573-82.
- Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
- Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
- Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
- Chen, Z.J. et al. (1996) Cell 84, 853-62.
- Senftleben, U. et al. (2001) Science 293, 1495-9.
- Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
- Xiao, G. et al. (2001) Mol Cell 7, 401-9.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
- 4886 Phospho-NF-KappaB p65 (Ser536) (93H1) Rabbit mAb (Alexa Fluor® 488 Conjugate)
- 3452 Rabbit IgG Isotype Control (Alexa Fluor® 647 Conjugate)
- 4764 NF-κB p65 (C22B4) Rabbit mAb
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
