WB, IP, IF-IC, FC-FP, ChIP, C&R
H M R
Endogenous
50 Active form. 120 Precursor
Rabbit IgG
#P25799
18033
Product Information
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:100 |
| Immunofluorescence (Immunocytochemistry) | 1:200 - 1:400 |
| Flow Cytometry (Fixed/Permeabilized) | 1:50 - 1:200 |
| Chromatin IP | 1:50 |
| CUT&RUN | 1:50 |
Storage
For a carrier free (BSA and azide free) version of this product see product #83394.
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ile415 of mouse NF-κB1 p105/p50 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).
Following IKK-mediated phosphorylation of p105 NF-κB at multiple sites (Ser921, 923, 927, and 932) on its carboxy terminus, SCF/β-TrCP-mediated processing produces the 50 kDa active form p50 (12,13).
- 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.
- Heissmeyer, V. et al. (2001) Mol Cell Biol 21, 1024-35.
- Orian, A. et al. (2000) EMBO J 19, 2580-91.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP C&R: CUT&RUN
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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