NF-kappaB p100/p52 ELISA Kit

Name: NF-kappaB p100/p52 ELISA Kit
SKU: DEIA-XYA1318
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Cell culture

Detection Method

iELISA

Intended Use

The NF-κB p100/p52 Cell-Based ELISA Kit is a convenient, lysatefree, high throughput and sensitive assay kit that can monitor NF-κB p100/p52 protein expression profile in cells. The kit can be used for measuring the relative amounts of NF-κB p100/p52 in cultured cells as well as screening for the effects that various treatments, inhibitors (ie. siRNA or chemicals), or activators have on NF-κB p100/p52.

Contents of Kit

Storage

Upon receipt, the kit should be stored at 4°C. The un-opened kit will be stable for up to 6 months from the date of shipment if stored at 4°C. Diluted Anti-NF-κB p100/p52 Antibody and diluted Anti-GAPDH Antibody can each be stored at 4°C for up to two weeks. HRP-Conjugated Anti-Rabbit IgG Antibody and HRP-Conjugated Anti-Mouse IgG Antibody will be stable at 4°C for up to six months. The SDS Solution should be stored at room temperature or warmed up to room temperature if stored at 4°C.

General Description

The Colorimetric Cell-Based ELISA Kit allows for the detection of various target proteins and the effects that certain stimulation conditions have on target protein expression in different cell lines. Qualitative determination of target protein concentration is achieved by an indirect ELISA format. In essence, the target protein is captured by target-specific primary (1°) antibodies while the HRP-conjugated secondary (2°) antibodies bind the Fc region of the 1° antibody. Through this binding, the HRP enzyme conjugated to the 2° antibody can catalyze a colorimetric reaction upon substrate addition. Due to the qualitative nature of the Cell Based ELISA, multiple normalization methods are described: 1) a monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target absorbance values. 2) Following the colorimetric measurement of HRP activity via substrate addition, the Crystal Violet whole-cell staining method is used to determine cell density. After staining, the results can be analyzed by normalizing the absorbance values to cell amounts, by which the plating difference can be adjusted. 3) If a phosphorylated target is being detected, an antibody against the non-phosphorylated counterpart will be provided for normalization purposes. The absorbance values obtained for the nonphosphorylated target can be used to normalize the absorbance values for the phosphorylated target.

Citations

Have you cited DEIA-XYA1318 in a publication? Let us know and earn a reward for your research.

Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Symptom Science: Advocating for Inclusion of Functional Genetic Polymorphisms

BIOLOGICAL RESEARCH FOR NURSING

Authors: Knisely, Mitchell R.; Maserati, Megan; Heinsberg, Lacey W.; Shah, Lisa L.; Li, Hongjin; Zhu, Yehui; Ma, Yumi; Graves, Letitia Y.; Merriman, John D.; Conley, Yvette P.

Abstract

Incorporating biologically based data into symptom science research can contribute substantially to understanding commonly experienced symptoms across chronic conditions. The purpose of this literature review was to identify functional polymorphisms associated with common symptoms (i.e., pain, sleep disturbance, fatigue, affective and cognitive symptoms) with the goal of identifying a parsimonious list of functional genetic polymorphisms with evidence to advocate for their inclusion in symptom science research. PubMed was searched to identify genes and functional polymorphisms associated with symptoms across chronic conditions, revealing eight functional genetic polymorphisms in seven different genes that showed evidence of association with at least three or more symptoms and/or symptom clusters: BDNF rs6265, COMT rs4680, FKBP5 rs3800373, IL-6 rs1800795, NFKB2 rs1056890, SLC6A4 5-HTTLPR+rs25531, and TNFA rs1799964 and rs1800629. Of these genes, three represent protein biomarkers previously identified as common data elements for symptom science research (BDNF, IL-6, and TNFA), and the polymorphisms in these genes identified through the search are known to impact secretion or level of transcription of these protein biomarkers. Inclusion of genotype data for polymorphisms offers great potential to further advance scientific knowledge of the biological basis of individual symptoms and symptom clusters across studies. Additionally, these polymorphisms have the potential to be used as targets to optimize precision health through the identification of individuals at risk for poor symptom experiences as well as the development of symptom management interventions.

Alteration of NF-kappa B activity leads to mitochondrial apoptosis after infection with pathological prion protein

CELLULAR MICROBIOLOGY

Authors: Bourteele, Soizic; Oesterle, Katja; Weinzierl, Andreas O.; Paxian, Stephan; Riemann, Marc; Schmid, Roland M.; Planz, Oliver

Abstract

Nuclear factor kappa B (NF-kappa B) is a key regulator of the immune response, but in almost the same manner it is involved in induction of inflammation, proliferation and regulation of apoptosis. In the central nervous system activated NF-kappa B plays a neuroprotective role. While in some neurodegenerative disorders the role of NF-kappa B is well characterized, there is poor knowledge on the role of NF-kappa B in prion disease. We found binding but no transcriptional activity of the transcription factor in vitro. Characterizing the mechanism of cell death after infection with pathological prion protein increased caspase-9 and caspase-3 activity was detected and the lack of NF-kappa B activity resulted in the inability to activate target genes that usually play an important role in neuroprotection. Additionally, we investigated the role of NF-kappa B after prion infection of Nfkb1(-/-), Nfkb2(-/-) and Bcl3(-/-) mice and central nervous system-specific p65-deleted mice revealing an accelerated prion disease in NF-kappa B2- and Bcl-3-deficient mice, which is in line with a reduced neuroprotective activity in prion infection. Based on our findings, we propose a model whereby the alteration of NF-kappa B activity at the early stages of infection with pathological prion protein leads to neuronal cell death mediated by mitochondrial apoptosis.