Description
Key facts
Detection method:Colorimetric,
Sample types:Nuclear Extracts,
Reacts with:Mouse, Rat, Human,
Assay type:Semi-quantitative,
Sensitivity:< 500 ng/well,
Assay time:3h 30m,
Assay Platform:Microplate reader
Product details:
C/EBP alpha/beta Transcription Factor Assay Kit (Colorimetric) (ab207199) is a high throughput assay to quantify C/EBP alpha/beta activation in nuclear extracts. This assay combines a quick ELISA format with a sensitive and specific non-radioactive assay for transcription factor activation.
A specific double stranded DNA sequence containing the C/EBP alpha/beta consensus binding site (5' –GCAAT– 3´) has been immobilized onto a 96-well plate. Active C/EBP alpha/beta present in the nuclear extract specifically binds to the oligonucleotide. C/EBP alpha/beta is detected by a primary antibody that recognizes an epitope of C/EBP alpha/beta accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout at OD 450 nm. This product detects human, mouse and rat C/EBP alpha/beta.
Key performance and benefits:
Assay time: 3.5 hours (cell extracts preparation not included).
Detection limit: < 0.5 μg nuclear extract/well.
Detection range: 0.5 – 10 μg nuclear extract/well.
C/EBPs (CCAAT-enhancer binding protein) basic leucine zipper transcription factors play an important role in regulating the balance between cell growth and differentiation. Expression of C/EBP is a function of tissue type as well as temporal period of development.
C/EBP beta is an important transcriptional activator involved in the regulation of several genes related to immune and inflammatory response and lipid storage. In addition, C/EBP beta is required for macrophage and B-cell development. The C/EBP beta regulator also plays a role in the IFN-signaling pathways and is a mediator of IL-6 signaling. C/EBP beta expression is enhanced by cytokines during acute phase reactions and may mediate the increase of acute phase proteins. In other systems, expression of C/EBP beta has been reported to be modulated by growth factor and C/EBP beta participates in growth hormone-regulated gene expression. Physiological and functional interactions of C/EBP with NFkB, Sp1 and other proteins have also been observed.
C/EBP alpha is a master regulator of myeloid progenitors, cooperates with other transcription factors and plays a critical role in early granulocyte development. Furthermore, C/EBP alpha is an inhibitor of cell proliferation, and is down-regulated in a large proportion of lung cancers.
There is a temporal order of expression for C/EBP alpha and C/EBP beta in differentiating cells, and C/EBP beta is often required for expression of C/EBP alpha. While C/EBP beta is induced early during adipocyte differentiation, C/EBP alpha is required in the late phase of adipocyte differentiation and up-regulates the battery of fat-specific genes required for the synthesis, uptake and storage of long chain fatty acids.
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
The 'CEBP Alpha' (also known as C/EBPα) and 'CEBP Beta' (also known as C/EBPβ) are members of the C/EBP family which belong to the basic leucine zipper class of transcription factors. These proteins function as critical regulators of gene expression. CEBPα has a molecular mass of approximately 42 kDa while CEBPβ is roughly 35 kDa to 38 kDa depending on isoform. C/EBPα is largely found in liver lung and adipose tissue while C/EBPβ shows expression in the liver adipose tissue and various myeloid cells.
Biological function summary
These transcription factors operate in the control of cell differentiation proliferation and metabolism. C/EBPα plays a vital role in the regulation of myeloid lineage commitment and functions synergistically with DNA-binding proteins to form a complex that influences adipogenesis and gluconeogenesis. C/EBPβ is similarly essential for granulopoiesis and lymphopoiesis making both important in immune function. They both engage in establishing and maintaining differentiated functions of certain cell types like adipocytes and hepatocytes after its initial expression.
Pathways
Both C/EBPα and C/EBPβ interact within essential metabolic and immune regulatory networks. They are part of the insulin signaling and cytokine receptor signaling pathways where they help modulate insulin-mediated effects on metabolism and control immune responses via cytokine regulation. In conjunction with other proteins like PPARγ and NF-κB C/EBP proteins help bridge metabolic and inflammatory pathways allowing adaptive responses to nutritional states.
C/EBPα is closely related to acute myeloid leukemia (AML) due to its role in hematopoietic differentiation where mutations impair normal blood cell development. C/EBPβ has been implicated in inflammatory diseases such as asthma due to its function in cytokine gene transcription. Both factors are interrelated in the context of disease with proteins such as GATA-1 and STAT3 which also contribute to these pathologies through similar pathways and regulatory mechanisms.
