Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.
Phospho-NF-κB p65 (Ser529) (E3K3J) Rabbit mAb #78764
Filter:
- WB
- IP
- IF
- F
Western blot analysis of extracts from THP-1 cells differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (50 ng/mL, 18 hr, and then rested 48 hr), untreated (-) or treated with Lipopolysaccharides (LPS) #14011 (5 μg/mL, 15 min; +), using Phospho-NF-κB p65 (Ser529) (E3K3J) Rabbit mAb (upper), NF-κB p65 (D14E12) XP® Rabbit mAb #8242 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Supporting Data
| REACTIVITY | H |
| SENSITIVITY | Endogenous |
| MW (kDa) | 65 |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IF-Immunofluorescence
- F-Flow Cytometry
Species Cross-Reactivity Key:
- H-Human
- Related Products
- Conjugates
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Fluorescent Western | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunofluorescence (Immunocytochemistry) | 1:3200 |
| Flow Cytometry (Fixed/Permeabilized) | 1:100 - 1:400 |
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 antibody.
For a carrier free (BSA and azide free) version of this product see product #61314.
For a carrier free (BSA and azide free) version of this product see product #61314.
Protocol
Specificity / Sensitivity
Phospho-NF-κB p65 (Ser529) (E3K3J) Rabbit mAb recognizes endogenous levels of NF-κB p65 protein only when phosphorylated at Ser529.
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser529 of human NF-κB p65 protein.
Background
Transcription factors of the nuclear factor κB (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 the 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 translocates to the nucleus (9-11).
Phosphorylation of NF-κB p65 by casein kinase II occurs following disassociation of NF-κB and IκB induced by cytokines, such as TNF-α (12).
Phosphorylation of NF-κB p65 by casein kinase II occurs following disassociation of NF-κB and IκB induced by cytokines, such as TNF-α (12).
- 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.
- Wang, D. et al. (2000) J Biol Chem 275, 32592-7.
Pathways
Explore pathways related to this product.
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