Product datasheet
C/EBP alpha/beta Transcription Factor Assay Kit
(Colorimetric) ab207199
2 Images
Overview
Product name
C/EBP alpha/beta Transcription Factor Assay Kit (Colorimetric)
Detection method
Colorimetric
Sample type
Nuclear Extracts
Assay type
Semi-quantitative
Sensitivity
< 500 ng/well
Assay time
3h 30m
Species reactivity
Reacts with: Mouse, Rat, Human
Product overview
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.
Notes
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
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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.
Tested applications
Suitable for: Functional Studies
Properties
Storage instructions
Please refer to protocols.
Components
1 x 96 tests
5 x 96 tests
10X Antibody Binding Buffer
1 x 2.2ml
1 x 11ml
10X Wash Buffer
1 x 22ml
1 x 110ml
96-well C/EBP assay plate
1 unit
5 units
Anti-rabbit HRP-conjugated IgG
1 x 11µl
1 x 55µl
Binding Buffer
1 x 10ml
1 x 50ml
C/EBP alpha beta antibody
1 x 11µl
1 x 55µl
Developing Solution
1 x 11ml
1 x 55ml
Dithiothreitol (DTT) (1 M)
1 x 100µl
1 x 500µl
Herring sperm DNA (1 μg/μL)
1 x 100µl
1 x 500µl
Lysis Buffer
1 x 10ml
1 x 50ml
Mutated oligonucleotide (20 pmol/µL)
1 x 100µl
1 x 500µl
Plate sealer
1 unit
5 units
Positive control nuclear extract (2.5 µg/µL)
1 x 40µl
1 x 200µl
Protease Inhibitor Cocktail
1 x 100µl
1 x 500µl
Stop Solution
1 x 11ml
1 x 55ml
Wild-type oligonucleotide (20 pmol/µL)
1 x 100µl
1 x 500µl
Cellular localization
CEBP Alpha: Nucleus. CEBP Beta: Nucleus.
Applications
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Our Abpromise guarantee covers the use of ab207199 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Functional Studies
Notes
Use at an assay dependent concentration.
C/EBP alpha/beta Transcription Factor Assay Kit (Colorimetric) images
Different amounts of unstimulated Jurkat
(grey) and U-937 (black) were tested for
C/EBP beta activation. These results are
provided for demonstration purposes only.
Different amounts of unstimulated Jurkat (Gray) and
U-937 (Black) were tested for C/EBP beta activation.
These results are provided for demonstration
purposes only.
Different amounts of unstimulated Jurkat
(grey) and U-937 (black) were tested for
C/EBP alpha activation. These results are
provided for demonstration purposes only.
Different amounts of unstimulated Jurkat (Gray) and
U-937 (Black) were tested for C/EBP alpha
activation. These results are provided for
demonstration purposes only.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Replacement or refund for products not performing as stated on the datasheet
Valid for 12 months from date of delivery
Response to your inquiry within 24 hours
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Extensive multi-media technical resources to help you
We investigate all quality concerns to ensure our products perform to the highest standards
If the product does not perform as described on this datasheet, we will offer a refund or replacement. For full details of the Abpromise,
please visit http://www.abcam.com/abpromise or contact our technical team.
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Terms and conditions
Guarantee only valid for products bought direct from Abcam or one of our authorized distributors
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