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NF-κB p65 (D14E12) XP®️ Rabbit mAb
synthetic peptide corresponding to residues surrounding Glu498 of human NF-κB p65/RelA protein
W, IP, IHC-P, IF-IC, F, ChIP
大量
CST
详见MSDS文件
详见说明书
H,M,R,Hm,Mk,Dg
单克隆
-20°c
pathway more info application references datasheet PDF MSDS PDF protocols
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 Dg=Dog
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
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W IP IHC-P IF-IC F ChIP | H M R Hm Mk Dg | Endogenous | 65 | Rabbit IgG |
Protocols |
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Specificity / Sensitivity | NF-κB p65 (D14E12) XP® Rabbit mAb recognizes endogenous levels of total NF-κB p65/RelA protein. It does not cross react with other NF-κB/Rel family members. |
Source / Purification | Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu498 of human NF-κB p65/RelA protein. Western BlottingWestern blot analysis of extracts from various cell lines using NF-κB p65 (D14E12) XP® Rabbit mAb. IHC-P (paraffin)Immunohistochemical analysis of paraffin-embedded human chronic cholecystitis using NF-κB p65 (D14E12) XP® Rabbit mAb. IHC-P (paraffin)Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or treated with hTNF-α #8902 (right), using NF-κB p65 (D14E12) XP® Rabbit mAb. Flow CytometryFlow cytometric analysis of HeLa cells using NF-κB p65 (D14E12) XP® Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). IF-ICConfocal immunofluorescent analysis of HT-1080 cells, untreated (left) or treated with hTNF-α #8902 (20 ng/ml, 20 min) (right), using NF-κB p65 (D14E12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Chromatin IPChromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells treated with hTNF-α #8902 (30 ng/ml, 1 hr) and either 5 μl of NF-κB p65 (D14E12) XP® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP® Human IκBα Promoter Primers #5552, human IL-8 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. |
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 is then translocated to the nucleus (9-11).
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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 |
For Research Use Only. Not For Use In Diagnostic Procedures. |
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