- 询价
- Cell Signaling Technology
- 6956
- USA
- 2025年12月11日
- W, IP, IHC-P, IF-IC, F, ChIP
- H,M,R,Hm,Mk,Mi,B,Dg,Pg
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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 抗体英文名：
NF-κB p65 (L8F6) Mouse mAb
- 抗原：
synthetic peptide corresponding to residues near the carboxy terminus of human NF-κB protein
- 应用范围：
W, IP, IHC-P, IF-IC, F, ChIP
- 保质期：
详见说明书
- 适应物种：
H,M,R,Hm,Mk,Mi,B,Dg,Pg
- 库存：
大量
- 供应商：
CST
- 级别：
详见MSDS文件
- 是否单克隆：
1
- 保存条件：
-20°c
- 规格：
100 ul (10 western blots)/carrier free & custom formulation / quantity
| 规格： | 产品价格： | ￥请询价 | |
|---|---|---|---|
| 规格： | 100 ul (10 western blots) | 产品价格： | ￥请询价 |
| 规格： | carrier free & custom formulation / quantity | 产品价格： | ￥请询价 |
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 Mi=Mink B=Bovine Dg=Dog Pg=Pig
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 |
|---|---|---|---|---|
| W IP IHC-P IF-IC F ChIP | H M R Hm Mk Mi B Dg Pg | Endogenous | 65 | Mouse IgG2b |
| Protocols |
|
|---|---|
| Specificity / Sensitivity | NF-κB p65 (L8F6) Mouse mAb recognizes endogenous levels of total NF-κB p65 protein. |
| Source / Purification | Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human NF-κB protein. Western Blotting
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® NF-κB p65 siRNA I #6261 (+), using NF-κB p65 (L8F6) Mouse mAb (upper) or α-Tubulin (11Η10) Rabbit mAb #2125 (lower). The NF-κB p65 (L8F6) Mouse mAb confirms silencing of NF-κB p65 expression, while the α-Tubulin (11Η10) Rabbit mAb is used as a loading control. Western Blotting
Western blot analysis of extracts from various cell lines using NF-κB p65 (L8F6) Mouse mAb. IHC-P (paraffin)
Immunohistochemical analysis of human chronic cholecystitis tissue using NF-κB p65 (L8F6) Mouse mAb. IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded OVCAR8 cell pellets treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (left) or treated with SignalSilence® NF-κB p65 siRNA I #6261 (right), using NF-κB p65 (L8F6) Mouse mAb. IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (right), using NF-κB p65 (L8F6) Mouse mAb. Flow Cytometry
Flow cytometric analysis of HeLa cells using NF-κB p65 (L8F6) Mouse mAb (blue) compared to a nonspecific negative control antibody (red). IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (20 ng/mL, 20 min; right), using NF-κB p65 (L8F6) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30 ng/ml, 1 hr) and either 10 μl of NF-κB p65 (L8F6) Mouse 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).
|
| 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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文献和实验【求助】测定NF-κB 活性参与者：sandy99实验中设计测定NF－κB 活性，我查的资料有以下几种选择：1、EMSA。请教所设计的标记探针的序列？以及该实验的注意事项。（没做过）2、抽提核蛋白，用P65抗体做Western Blot.请问性价比较高的，好做的是哪个公司的抗体？还有啊，如果这个抗体用FITC标记过，是不是就可以用来做激光共聚焦实验？也可做WB？3、ELISA，贵，暂不考虑。谢谢！参与者：hwclove我试着说一下我的看法：1.EMSA中一般都是用P标记的探针，具有放射
大鼠核因子-κB亚基p65亲和肽(NF-κB p65)酶联免疫分析
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gmol／L的培养基培养Hela细胞4h后弃培养基并以无热原生理盐水漂洗细胞三遍，再加入含TNFα 20ng／ml的培养基继续培养。③空白对照组：培养基为不含血清，不加氯化镧和TNFα。④氯化镧对照组：以含氯化镧100gmol／L的培养基培养Hela细胞。各组细胞根据检测方法的要求分别培养于24孔培养板或培养瓶中，于30min后，收集培养上清、细胞待检。运用Westernblot技术测定胞浆提取物中IrBa蛋白表达和磷酸化水平及胞核提取物中NF-κB／p65蛋白表达水平；最后用Transcription
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