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PathScan® Sandwich ELISA Kit

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REACTIVITY
Product Includes Volume Solution Color
Phospho-NF-κB p65 (Ser536) Mouse mAb coated microwells 96 tests
NF-κB p65 Rabbit Detection mAb 11 milliliter equivalent Green (Lyophilized)
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 1 Ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Sealing Tape 2 sheets
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

Product Description

CST's PathScan® Phospho-NF-KappaB p65 (Ser536) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-NF-KappaB p65 protein. A Phospho-NF-KappaB p65 (Ser 536) Mouse mAb has been coated onto the microwells. After incubation with cell lysates, phospho-NF-KappaB p65 protein is captured by the coated antibody. Following extensive washing, NF-KappaB p65 Rabbit mAb is added to detect the captured phospho-NF-KappaB p65 protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho-NF-KappaB p65 (Ser536).

Antibodies in kit are custom formulations specific to kit.


Specificity / Sensitivity

CST's PathScan® Phospho-NF-KappaB p65 (Ser536) Sandwich ELISA Kit detects endogenous levels of phospho-NF-KappaB p65 at Ser536. As shown in Figure 1, using the Phospho-NF-KappaB p65 (Ser536) ELISA Kit #7173, a significant induction of Phospho-NF-KappaB p65 (Ser536) in HeLa cells treated with TNF-alpha is detected. However, levels of NF-KappaB p65 (either untreatead or treated), detected by the Total NF-KappaB p65 Sandwich ELISA Kit #7174, remain unchanged. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.


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).


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.

12.  Peiser, M. et al. (2008) J Leukoc Biol , .


Entrez-Gene Id 5970
Swiss-Prot Acc. Q04206


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
PathScan® is a trademark of Cell Signaling Technology, Inc.