Product Pathways - PathScan ELISA
PathScan® Total NF-κB p65 Sandwich ELISA Antibody Pair #7836
|Kit Includes||Volume||Cap Color|
|NF-kB p65 Rabbit Capture Ab (100X)||0.4 ml||Pink|
|NF-kB p65 Mouse Detection Ab (100X)||0.4 ml||Blue|
|Anti-mouse IgG, HRP-linked Antibody||0.04 ml||Yellow|
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
CST's PathScan® Total NF-κB p65 Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total NF-κB p65 Sandwich ELISA Kit #7174. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The NF-κB p65 Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a NF-κB p65 Detection Antibody and anti-Mouse IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of total NF-κB p65 protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The relationship between lysate protein concentration from untreated and TNF-α and IL-1β treated HeLa cells and the absorbance at 450 nm using PathScan® Total NF-κB p65 Sandwich ELISA Antibody Pair #7836 is shown. HeLa cells were treated with TNF-α and IL-1β for 5 minutes at 37ºC and then lysed.
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).
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- 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.
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- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
- 9803 Cell Lysis Buffer (10X)
- 7174 PathScan® Total NF-κB p65 Sandwich ELISA Kit
- 7173 PathScan® Phospho-NF-κB p65 (Ser536) Sandwich ELISA Kit
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 8242 NF-κB p65 (D14E12) XP® Rabbit mAb
- 4764 NF-κB p65 (C22B4) Rabbit mAb
For Research Use Only. Not For Use In Diagnostic Procedures.