COP1 Antibody
Description
Introduction to COP1 Antibody
COP1 (Constitutive Photomorphogenic 1) antibodies are specialized immunological tools designed to detect and study the COP1 protein, an E3 ubiquitin ligase encoded by the RFWD2 gene in humans. COP1 regulates critical cellular processes, including protein degradation, cell cycle progression, and stress responses, by targeting substrates like FoxO1, p27, and c-Jun for ubiquitination . These antibodies enable researchers to investigate COP1's expression, localization, and functional roles in both physiological and pathological contexts, such as cancer metabolism and photomorphogenesis .
Applications in Research
COP1 antibodies are widely used in molecular biology and clinical research. Key applications include:
Novus Biologicals (NBP2-92699):
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Host Species: Rabbit
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Reactivity: Human, mouse, rat
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Dilution Range: 1:500–1:2000 (WB)
Proteintech (13542-1-AP):
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Host Species: Rabbit
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Reactivities: Human, mouse, rat
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Dilution Range: 1:50–1:500 (IHC), 1:500–1:1000 (WB)
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Observed Molecular Weight: 90 kDa (vs. calculated 80 kDa due to post-translational modifications) .
Validation Data:
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Western Blot: Confirmed in human heart tissue and mouse models .
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IHC: Strong staining in poorly differentiated gastric cancer tissues compared to normal tissues .
Mechanistic Insights:
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FoxO1 Degradation: COP1 promotes ubiquitination and proteasomal degradation of FoxO1, suppressing gluconeogenic genes like G6Pase and PEPCK in hepatoma cells .
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Cell Cycle Regulation: COP1 interacts with p27 during G1 phase, accelerating its degradation to drive cell proliferation .
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Cancer Metastasis: In gastric cancer, COP1 overexpression degrades CDH18, activating the PI3K/AKT pathway to enhance tumorigenesis .
COP1 in Cancer:
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Gastric Cancer: High COP1 expression correlates with advanced tumor infiltration (T stage) and poor prognosis .
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TNBC and Colorectal Cancer: Cop1 knockout in murine models reduces tumor growth and enhances anti-PD-1 therapy efficacy .
Therapeutic Targeting:
COP1’s dual role as an oncoprotein (degrading tumor suppressors like p27) and tumor suppressor (targeting oncoproteins like c-Jun) highlights its potential as a cancer therapy target .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- COL12 is a substrate of COP1. PMID: 29187570
- COP1 plays a significant role in drought stress tolerance. PMID: 30139551
- Mutation of the coiled-coil domain in COP1, which prevents dimer formation, impairs its function in coordinating flowering time. PMID: 29273730
- Nitrate Reductase levels are negatively regulated by COP1 and ammonium. PMID: 29662028
- Data suggests that light directly interacts with the inducer of CBF Expression (ICE) transcription factors (ICE)-directed signaling module, via the COP1-mediated protein surveillance system, in the modulation of stomatal development. PMID: 29070509
- The COP1/SPA complex associates with and stabilizes PHYTOCHROME INTERACTING FACTOR 3 (PIF3) to repress photomorphogenesis in the dark. PMID: 28292892
- High COP1 expression is linked to defense against turnip crinkle virus. PMID: 29513740
- COP1 mediates the dark-specific degradation of microtubule-associated protein WDL3 in regulating Arabidopsis hypocotyl elongation. PMID: 29087315
- Thermal activation of COP1 enables coincidence between warm temperature signaling and circadian rhythms, allowing plants to time hypocotyl thermomorphogenesis optimally at warm temperatures. PMID: 28418582
- ABI4 and HY5 antagonistically regulate the expression of COP1 and the subsequent greening process. Conversely, ABI4 and HY5 are targeted for degradation by COP1 in the light and dark, respectively, ensuring a balanced interplay between their actions during seedling de-etiolation. PMID: 27255835
- Data indicates the localization of UVR8 signaling in the nucleus and a dual role for COP1 in the regulation of UV-B-induced UVR8 nuclear accumulation and in UVR8-mediated UV-B signaling. PMID: 27407149
- SPA proteins have a dual role: (1) they are essential for light-responsiveness of COP1 subcellular localization, and (2) they enhance COP1 activity in darkness independent of COP1 nuclear import/retention. PMID: 28536102
- The alpha2-cop mutant exhibited defects in plant growth, including small rosettes, stems, and roots, and mislocalization of p24delta5, a protein involved in COPI binding. The mutant also displayed abnormal Golgi apparatus morphology. PMID: 28025315
- Exposure to blue light is necessary for an in vivo association of CRY1 and CRY2 with COP1. PMID: 28991901
- DHU1 negatively regulates UV-B signaling through its direct interaction with COP1 and RUP1 (At5g52250). PMID: 28735869
- BBX21 is a crucial component involved in the COP1-HY5 regulatory hub. PMID: 27325768
- Transcript levels of ABA biosynthesis genes are higher in cip1-1 than in the wild-type, suggesting that CIP1 is positively involved in ABA response. PMID: 27372427
- Salt stress and ethylene antagonistically regulate the nucleocytoplasmic partitioning of COP1. PMID: 26850275
- Genetic and biochemical studies identify a function for SIZ1 in photomorphogenesis and reveal a novel SUMO-regulated ubiquitin ligase, COP1, in plants. PMID: 27128446
- Data suggests a coordinated regulation of Arabidopsis proteins SHW1, COP1, and HY5 in seedling development. PMID: 26474641
- COP1 plays a role in CONSTANS protein degradation during photoperiodic flowering. PMID: 26358558
- CSU2 interacts with COP1 via the coiled-coil domain association. CSU2 negatively regulates COP1 E3 ubiquitin ligase activity. PMID: 26714275
- It is proposed that light perceived by phytochromes causes a switch in the ubiquitination activity of COP1/SPA2 from ubiquitinating downstream substrates to ubiquitinating SPA2, subsequently leading to a repression of COP1/SPA2 function. PMID: 26368289
- CUL4(COP1-SPA) E3 ubiquitin ligase is necessary for the light-induced degradation of PIF1 in Arabidopsis. PMID: 26037329
- Genetic analyses with transgenes expressing a genomic pmARI12:ARI12-GFP construct confirm the epistatic interaction between COP1 and ARI12 in growth responses to high fluence rate UV-B. PMID: 25817546
- AN3 may act with other proteins that bind to the COP1 promoter to promote anthocyanin accumulation and inhibit light-induced root elongation. PMID: 25256341
- Two distinct domains of the UVR8 photoreceptor interact with COP1 to initiate UV-B signaling in Arabidopsis. PMID: 25627067
- Cytoplasmic partitioning of COP1 under light is essential to protect HYL1 against protease X. PMID: 25532508
- Mutations in the key repressor of light signaling, the COP1/SPA complex, cause a strong hyperaccumulation of anthocyanins under both normal and high light conditions. PMID: 25482806
- Light and COP1 regulate the level of overexpressed DET1 protein. PMID: 25575996
- Analysis of BBX22 degradation kinetics shows that it has a short half-life under both dark and light conditions. COP1 mediates the BBX22 degradation in the dark. While dispensable in the dark, HY5 contributes to the BBX22 degradation in the light. PMID: 21427283
- COP1 is identified as a potential coordinator of cytoskeletal and electrophysiological activities required for guard cell function. PMID: 25151660
- Molecular and biochemical evidence suggests that the UVR8-COP1 affinity in plants is crucial for determining the photomorphogenic UV-B signal transduction coupling with UVR8-mediated UV-B light perception. PMID: 24651064
- COP1 inactivation involved in rapid light-induced responses was compared to that of nuclear HY5. PMID: 24434030
- Research demonstrates that the COP1-HY5 complex is a novel integrator that plays a vital role in ethylene-promoted hypocotyl growth in the light. PMID: 24348273
- The MID protein contributes to COP1/SPA1-controlled repression of flowering under short-day conditions. PMID: 23857347
- COP1 plays a role in establishing the daily patterns of sensitivity to shade in the fluctuating light environments of plant canopies. PMID: 23647163
- A functional connection between COP1 and TOPOVI in plants links COP1-dependent development with the regulation of endoreduplication. PMID: 23573936
- The COP1/SPA complex affects PAP1 and PAP2 both transcriptionally and post-translationally. This complex controls anthocyanin levels in Arabidopsis. PMID: 23425305
- UV-B-induced reorganization of COP1 complexes achieves a functional switch of COP1 from repressing to promoting photomorphogenesis. PMID: 24067658
- The light-induced decline of phyA levels is reduced in spa mutants regardless of the growth medium, suggesting a COP1-independent role for SUppressor of phytochrome A proteins. PMID: 23391578
- In cop1 mutants, ddb1b-2 enhanced the cop1-4 short hypocotyl phenotype in both dark and light, increased anthocyanin levels in cop1-1 under light conditions, but had no effect in adult plants. PMID: 23450167
- Alanine mutants of specific tryptophan residues appear monomeric and constitutively bind COP1 in plants, but their responses indicate that monomer formation and COP1 binding alone are not sufficient for UVR8 function. PMID: 23012433
- COP1 gene expression in response to photomorphogenic UV-B is controlled by a combinatorial regulation of FHY3 and HY5. This UV-B-specific working mode of FHY3 and HY5 is distinct from their roles in far-red light and circadian conditions. PMID: 23150635
- COP1/SPA activity, via LONG HYPOCOTYL IN FR LIGHT1, is required for shade-induced modulation of the auxin biosynthesis pathway, enhancing cell elongation in low red:far red light conditions. PMID: 23093358
- Ethylene-promoting hypocotyl growth via IAA is mediated by light, and COP1 significantly influences the transcription of genes downstream of EIN3. Therefore, COP1 plays a crucial role in the contrasting effects of ethylene on hypocotyl elongation. PMID: 22890836
- A Mediator component collaborates with COP1 in regulating light responses, and the hypersensitive seedling phenotype is strictly dependent on the presence of HY5, a key positive regulator of light-dependent gene expression. PMID: 22760208
- COP1 mediates the coordination of root and shoot growth by light through modulation of PIN1- and PIN2-dependent auxin transport in Arabidopsis. PMID: 22912415
- COP1 interacts with BBX24 in vivo in a UV-B-dependent manner, requiring UV-B-induced COP1 accumulation. PMID: 22410790
- Mutations in the region responsible for the interaction with COP1 indicate that a physical interaction of the proteins is also necessary for degradation of BBX24 in the light and for normal photomorphogenesis. PMID: 21685177
Q&A
Experimental Design: Choosing the Right COP1 Antibody
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Question: What factors should I consider when selecting a COP1 antibody for my research?
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Answer: When choosing a COP1 antibody, consider the species reactivity, application suitability (e.g., Western Blot, Immunoprecipitation, Immunohistochemistry), and the immunogen used. For example, Abcam's ab70890 is a rabbit polyclonal antibody suitable for human samples and immunoprecipitation , while ab56400 is a mouse monoclonal antibody suitable for multiple applications including flow cytometry and reacts with both human and mouse samples .
Data Interpretation: Understanding COP1 Function
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Question: How does COP1 function as an E3 ubiquitin ligase, and what are its implications in cellular processes?
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Answer: COP1 acts as an E3 ubiquitin ligase by mediating the ubiquitination and subsequent proteasomal degradation of target proteins. It plays roles in regulating proteins like FoxO1, MTA1, and p53, impacting cell survival, metabolism, and cancer progression . Understanding these functions helps in interpreting data related to COP1's role in cellular processes.
Methodological Considerations: Immunoprecipitation and Western Blot
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Question: What are the optimal conditions for using COP1 antibodies in immunoprecipitation (IP) followed by Western Blot (WB)?
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Answer: For IP, use a suitable antibody concentration (e.g., 3 µg/mL for ab70890) and ensure sufficient protein input (e.g., 1 mg for IP). For WB, use a lower antibody concentration (e.g., 1 µg/mL) and optimize detection conditions (e.g., chemiluminescence with a short exposure time) . Ensure proper controls, such as IgG controls, to validate specificity.
Advanced Research: Analyzing COP1 Interactions
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Question: How can I investigate the interactions between COP1 and its target proteins using co-immunoprecipitation assays?
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Answer: To study COP1 interactions, perform co-immunoprecipitation assays using tagged proteins (e.g., HA-tagged FoxO1 and FLAG-tagged COP1). Pretreat cells with proteasome inhibitors (e.g., ALLN) to stabilize interactions. Analyze immunoprecipitates by Western Blot to detect co-precipitated proteins .
Contradictory Data Analysis: Understanding COP1's Role in Different Systems
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Question: How do I reconcile contradictory findings regarding COP1's role in different biological systems (e.g., plant vs. mammalian systems)?
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Answer: Consider the context-specific functions of COP1. In plants, COP1 is involved in photomorphogenesis and pathogen resistance , while in mammals, it regulates cell survival and metabolism . Analyze experimental conditions, species-specific differences, and potential post-translational modifications that might influence COP1's activity.
Methodological Optimization: Enhancing COP1 Antibody Specificity
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Question: What strategies can I use to enhance the specificity of COP1 antibodies in complex biological samples?
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Answer: To enhance specificity, use high-quality antibodies with well-characterized immunogens, optimize antibody concentrations, and include appropriate controls (e.g., IgG controls). Consider using peptide competition assays to validate specificity .
Advanced Techniques: Investigating COP1-Mediated Ubiquitination
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Question: How can I investigate COP1-mediated ubiquitination of target proteins?
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Answer: To study COP1-mediated ubiquitination, co-transfect cells with tagged ubiquitin and COP1 expression vectors. Use immunoprecipitation followed by Western Blot to detect ubiquitinated proteins. Treat cells with proteasome inhibitors to stabilize ubiquitinated species .
Data Integration: Combining COP1 Studies Across Different Species
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Question: How can I integrate findings from COP1 studies across different species to understand its conserved functions?
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Answer: Compare the structural and functional conservation of COP1 across species. Analyze the role of COP1 in similar biological processes (e.g., protein degradation pathways) and consider species-specific adaptations. Use bioinformatics tools to align sequences and predict functional motifs .
Experimental Validation: Confirming COP1 Interactions
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Question: What methods can I use to validate the interactions between COP1 and other proteins in vivo?
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Answer: Validate COP1 interactions using co-immunoprecipitation assays in vivo. Employ techniques like GST pull-down assays to assess direct interactions. Additionally, use cellular fractionation to study the subcellular localization of interacting proteins .
