NF-κB Family Member Antibody Sampler Kit #4766
Product Information
Kit Usage Information
Protocols
- 3017: Western Blotting, Immunohistochemistry (Paraffin), Flow, CUT&RUN Assay, CUT&Tag
- 4717: Western Blotting, Immunoprecipitation (Magnetic)
- 4882: Western Blotting, Immunoprecipitation (Agarose)
- 4922: Western Blotting, Immunoprecipitation (Agarose), CUT&RUN Assay
- 6956: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow, ChIP Magnetic
- 7074: Western Blotting
- 7076: Western Blotting
- 8242: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow, ChIP Magnetic, Chromatin IP-seq, CUT&RUN Assay
- 12540: Western Blotting, Immunoprecipitation (Magnetic), Immunohistochemistry (Paraffin), ChIP Magnetic
- 12707: Western Blotting
Product 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).
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 (D7H5M) Rabbit mAb #12540 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.
Source / Purification
Monoclonal antibodies are 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, surrounding Leu65 of human c-Rel protein, surrounding Gly415 of human NF-κB p105/p50 protein, surrounding Glu498 of human NF-κB p65/RelA protein, and near the amino terminus of human NF-κB2 p100/p52. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino acid residues at the the carboxy terminus of human NF-κB1 p105 and near the amino terminus of human NF-κB2 p100/p52. Polyclonal antibodies are purified by Protein A and peptide affinity chromatography.
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).
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