Expedeon/ELISA-ONE™ for NF-κB p65/ELISA-ONE for total NF-κB p65, 1 x 96 Well Plate Kit/ELK008

ELISA-ONE for Phospho NF-κB p65 (pS536)

Specificity

ELISA-ONE for phospho NF-κB p65 (pS536) are ready to use ELISA kits for the detection of endogenous levels of NF-κB p65 when phosphorylated at Ser536.

ELISA-ONE for phospho NF-κB p65 (pS536) is comprised of a precoated 96 well (12×8 strips) plate and an antibody pair, recognizing the phospho-Ser536 epitope and a distal epitope of NF-κB p65.

The antibody pair recognizes human and mouse NF-κB p65. However, other species should be tested on a case by case basis.

ELISA-ONE for Total NF-κB p65

Specificity

ELISA-ONE for total NF-κB p65 are ready to use ELISA kits for the detection of endogenous levels of NF-κB p65 irrespective of phosphorylation status.

ELISA-ONE for total NF-κB p65 is comprised of a precoated 96 well (12×8 strips) plate and an antibody pair, recognizing two invariants epitopes of NF-κB p65.

The antibody pair recognizes human and mouse NF-κB p65. However, other species should be tested on a case by case basis.

ELISA-ONE for Phospho & Total NF-κB p65

Specificity

ELISA-ONE for phospho and total NF-κB p65 are ready to use ELISA kits for the detection of endogenous levels of NF-κB p65 in its phosphorylated and unphosphorylated form.

ELISA-ONE for phospho and total NF-κB p65 is comprised of a precoated 96 well (12×8 strips) plate and two antibody pairs, recognizing the phospho-Ser536 epitope and an invariant epitope of NF-κB p65.

The antibody pair recognizes human and mouse NF-κB p65. However, other species should be tested on a case by case basis.

Technical Specifications

Species Cross-reactivity

Tested: Human, mouse

Kit Contents

• 1x 96 well (12×8 strips) CaptSure assay plate
• 1x adherent plate seal
• 1x Lysis Buffer (5X)
• 1x Enhancer Solution
• 1x Wash Buffer (10X)
• 1x Detection Reagent
• 1x Stop Solution
• 1x Lyophilized Positive Control Lysate
• 1x Capture Antibody Reagent 1
• 1x Capture Antibody Reagent 2 (phospho / total target kits only)
• 1x Detection Antibody Reagent 1
• 1x Detection Antibody Reagent 2 (phospho / total target kits only).

NF-κB p65 Signaling Pathway

The nuclear factor κB transcription factor family plays a pivotal role in inflammatory and immune responses. There are five family known members, namely p65 (RelA), c‐Rel, RelB, NF‐κB1 (p105/p50) and NF‐κB2 (p100/p52). The p50 and p52 products form dimeric complexes with Rel proteins, which bind DNA and regulate transcription. The most abundant form in many cell types is the p65/p50 complex. In unstimulated cells, NF‐κB is inactive in the cytoplasm, bound by I‐κB proteins. Activation of the NF‐κB pathway induces phosphorylation of I‐κBs, targeting them for degradation through the ubiquitin proteasome pathway. Free NF‐κB can then enter the nucleus, where it regulates expression of its target genes.

NF-κB p65 References

Buss, H., Handschick, K., Jurrmann, N., Pekkonen, P., Beuerlein, K., Müller, H., Wait, R., Saklatvala, J., Ojala, PM., Schmitz, ML., Naumann, M., Kracht. M. Cyclin-dependent kinase 6 phosphorylates NF-κB P65 at serine 536 and contributes to the regulation of inflammatory gene expression. PLoS One. 2012;7(12):e51847. doi: 10.1371/journal.pone.0051847. Epub 2012 Dec 26.

Pan, X., Jiang, B., Liu, J., Ding, J., Li, Y., Sun, R., Peng, L., Qin, C., Fang, S., Li, G. STC1 promotes cell apoptosis via NF-κB phospho-P65 Ser536in cervical cancer cells. Oncotarget. 2017 Jul 11;8(28):46249–46261. doi: 10.18632/oncotarget.17641.

Zhang, L., Shao, L., Creighton, CJ., Zhang, Y., Xin, L., Ittmann, M., Wang, J. Function of phosphorylation of NF-kB p65 ser536 in prostate cancer oncogenesis. Oncotarget. 2015 Mar 20;6(8):6281–4.

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