Product Pathways - NF-kB Signaling

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NF-κB Family Member Antibody Sampler Kit #4766

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
NF-κB p65 (L8F6) Mouse mAb #6956	40 µl	W IP IHC-P IF-IC F ChIP	H M R Hm Mk Mi B Dg Pg	65	Mouse IgG2b
RelB (C1E4) Rabbit mAb #4922	40 µl	W IP	H M R Mk	70	Rabbit IgG
c-Rel Antibody #4727	40 µl	W IP IHC-P IF-IC F	H Mk	78	Rabbit
NF-κB1 p105/p50 Antibody #3035	40 µl	W IP ChIP	H Mk	50 Active form. 120 Precursor.	Rabbit
NF-κB1 p105 Antibody #4717	40 µl	W IP	H M R Mk Mi B Pg	120	Rabbit
NF-κB2 p100/p52 Antibody #4882	40 µl	W IP	H M R Mk	52 (mature). 120 (precursor).	Rabbit
NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific) #3017	40 µl	W IHC-P F	H Mk	52 active form. 120 precursor.	Rabbit IgG
NF-κB p65 (D14E12) XP® Rabbit mAb #8242	40 µl	W IP IHC-P IF-IC F ChIP	H M R Hm Mk Dg	65	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat
Anti-mouse IgG, HRP-linked Antibody #7076	100 µl				Horse

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Mi=Mink  B=Bovine  Dg=Dog  Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in this kit recognizes endogenous levels of its target protein regardless of post-translational modification state such as phosphorylation or acetylation. The NF-κB1 p105/p50 Antibody #3035 detects both the precursor protein p105 and its cleavage product p50, while the NF-κB1 p105 Antibody #4717 only detects p105 and will not cross-react with p50. Both the NF-κB2 p100/p52 Antibody #4882 and the NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific) #3017 will cross-react with the precursor protein p100 and its cleavage product p52.

Description

This kit contains reagents to examine total protein levels of the five NF-κB/Rel family members: p65/RelA, RelB, c-Rel, NF-κB1 (p105/p50) and NF-κB2 (p100/p52).

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino acid residues surrounding Glu498 of human NF-κB p65, near the carboxy terminus of human c-Rel, near the amino terminus of human NF-κB1 p105/p50, at the carboxy terminus of human NF-κB1 p105 and near the amino terminus of human NF-κB2 p100/p52. Antibodies are purified by Protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to amino acid residues near the carboxy terminus of human NF-κB p65, surrounding Ser424 of human RelB and near the amino terminus of human NF-κB2 p100/p52.

Background

Transcription factors of the nuclear factor κ 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 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).

1. Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
2. Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
3. Haskill, S. et al. (1991) Cell 65, 1281-9.
4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
7. Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
8. Chen, Z.J. et al. (1996) Cell 84, 853-62.
9. Senftleben, U. et al. (2001) Science 293, 1495-9.
10. Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
11. 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!

Protocols

3017:

Flow, IHC / Paraffin, Western Blotting

3035:

ChIP Agarose, ChIP Magnetic, Immunoprecipitation, Western Blotting

4717:

Immunoprecipitation, Western Blotting

4727:

Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

4882:

Immunoprecipitation, Western Blotting

4922:

Immunoprecipitation, Western Blotting

6956:

ChIP Agarose, ChIP Magnetic, Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

8242:

ChIP Agarose, ChIP Magnetic, Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

Companion Products

• 9936 NF-κB Pathway Sampler Kit
• 3039 Phospho-NF-κB p65 (Ser468) Antibody
• 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
• 4806 Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb
• 4808 Phospho-NF-κB p105 (Ser933) (178F3) Rabbit mAb (IHC Specific)
• 4810 Phospho-NF-κB2 p100 (Ser866/870) Antibody
• 9243 NF-κB Control Cell Extracts
• 7071 Phototope^®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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For Research Use Only. Not For Use In Diagnostic Procedures.