DEK Recombinant Monoclonal Antibody
Description
Overview of DEK Recombinant Monoclonal Antibody
DEK recombinant monoclonal antibodies are laboratory-engineered immunoglobulins designed to specifically target the DEK oncoprotein, a nuclear protein implicated in transcriptional regulation, chromatin remodeling, and cancer progression . These antibodies are produced using recombinant DNA technology, ensuring precise sequence definition, batch-to-batch consistency, and high specificity . DEK (Gene ID: 7913) is associated with leukemias, solid tumors, and autoimmune diseases, making it a critical target for diagnostics and therapeutic research .
Applications in Research and Diagnostics
DEK recombinant monoclonal antibodies are pivotal in:
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Western Blot (WB): Detects DEK at ~50 kDa in nuclear extracts (e.g., HeLa, Jurkat cells) .
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Immunohistochemistry (IHC): Localizes DEK in nuclei of human cervical cancer tissues at dilutions up to 1:4,000 .
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Immunofluorescence (IF): Visualizes DEK in HepG2 and K562 cell lines with minimal cross-reactivity .
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Flow Cytometry (FC): Identifies DEK-expressing cells in Sézary syndrome PBMCs (21% TRBV5-1+ cells) .
Research Findings and Case Studies
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Oncogenic Role: DEK overexpression correlates with poor prognosis in hepatocellular carcinoma and promotes tumor cell survival .
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Therapeutic Potential: Recombinant DEK antibodies enable targeted depletion of malignant T cells (e.g., TRBV5-1+ clones) with minimal off-target effects .
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Mechanistic Insights: DEK regulates hematopoietic stem cell quiescence via the NCoR1/HDAC3-Akt/mTOR axis, as shown in siRNA knockdown studies .
Figure 1: DEK Antibody Validation Data
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Western Blot: Distinct ~50 kDa band in nuclear extracts of HeLa and Jurkat cells .
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IHC: Strong nuclear staining in human cervical cancer tissues (Bio-Techne MAB7020) .
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Flow Cytometry: Specific binding to DEK+ T cells (ab221545, Abcam) .
Comparative Advantages Over Traditional Antibodies
Future Directions
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Clinical Translation: Defucosylated DEK antibodies (e.g., akin to Mogamulizumab) could enhance antibody-dependent cellular cytotoxicity (ADCC) in leukemia therapies .
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Multi-Omics Integration: Coupling DEK antibodies with spatial transcriptomics (e.g., RNAscope™) for tumor microenvironment analysis .
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Open-Source Repositories: Platforms like the ABCD database enable sharing of sequenced DEK antibodies to accelerate global research .
Product Specs
CUSABIO developed a recombinant monoclonal antibody against DEK by immunizing a rabbit with a synthesized peptide representing human DEK. Following immunization, B cells were isolated from the rabbit, and RNA was extracted. This RNA was reverse-transcribed into cDNA, which served as a template for extending DEK antibody genes using degenerate primers. These extended DEK antibody genes were then integrated into a plasmid vector and introduced into host cells for expression. The DEK recombinant monoclonal antibody was purified from the cell culture supernatant through affinity chromatography. It has been validated for its effectiveness in ELISA, IHC, IF, and FC applications, demonstrating specificity for human DEK protein.
DEK, a nuclear protein, has been implicated in both normal cellular functions and disease-related processes. It is known to play a crucial role in various cellular processes within the cell nucleus, including chromatin organization, DNA binding, transcription regulation, cell cycle regulation, and immune response.
Target Background
DEK is involved in chromatin organization.
- This research identifies DEK as a critical and multifaceted mediator of homologous recombination. PMID: 28317934
- DEK is detected in spontaneously forming neutrophil extracellular traps (NETs) from juvenile idiopathic arthritis patient synovial neutrophils, and DEK-targeted aptamers reduce NETs formation. PMID: 28165452
- High DEK proto-oncogene protein (DEK) expression is associated with a complete response to neoadjuvant chemoradiotherapy. PMID: 29409457
- Elevated DEK expression is linked to cervical cancer tumorigenesis and metastasis. PMID: 28627610
- The findings of this study suggest that epithelial-mesenchymal transition (EMT) of colorectal carcinoma cells is partially mediated by DEK, potentially impacting the invasive capacity of these cells. Additionally, cell proliferation and apoptosis were susceptible to DEK silencing. PMID: 29115492
- RNA sequencing proved to be a valuable tool for identifying a fusion of genes DEK and NUP214 in a leukemia displaying a cryptic cytogenetic rearrangement of chromosome band 9q34. PMID: 29109093
- Overexpression of DEK enhances carcinogenesis, including field cancerization, in oral squamous cell carcinoma (OSCC) by stimulating the G1/S phase transition and promoting DNA replication. PMID: 28834425
- DEK and LCMR1 were shown to cooperate in inhibiting apoptosis in lung cancer cells. PMID: 28765911
- High DEK expression is associated with breast cancer angiogenesis. PMID: 26988756
- Our study indicates that DEK expression is essential for tumorigenesis and metastasis of hepatocellular carcinoma. PMID: 27057626
- High DEK expression is linked to melanoma progression. PMID: 27893188
- DEK correlates with IL6 expression in HPV16+/p16+ oropharyngeal squamous cell carcinomas (OPSCC). PMID: 28423581
- Data support a scenario where overexpression of the human DEK oncogene reprograms keratinocyte metabolism to meet the energy and macromolecule demands required for sustaining cancer cell growth. PMID: 28558019
- Decreased DEK expression in plasma cells suggests a potential role of this gene in plasma cell development, and the lack of detectable DEK protein by immunohistochemistry (IHC) could serve as a biomarker for normal and malignant plasma cells. PMID: 28558048
- Our data suggest that DEK plays multiple roles in facilitating tumor growth and maintenance. It can be used as a potential target for astrocytic tumor diagnosis and gene therapy. PMID: 28670979
- High DEK expression is associated with pancreatic ductal adenocarcinoma. PMID: 27959420
- AP-2a is a significant transcription factor of DEK expression, which is correlated with the methylation level of the DEK core promoter in hepatocellular carcinoma. PMID: 27499261
- miR-592 targets DEK transcript and suppresses hepatocellular carcinoma cell growth. PMID: 26722432
- Results indicate that DEK binds to transcription start sites with a dual role in activation and repression of highly and ubiquitously expressed genes. PMID: 25216995
- The study identified the DEK oncoprotein as a critical factor that interacts with an essential upstream enhancer element of the EKLF promoter and exerts a positive effect on EKLF levels. PMID: 26303528
- DEK promotes the growth of colorectal cancer and apoptosis. PMID: 25340858
- Data show that oncoprotein DEK facilitates cellular proliferation under conditions of DNA replication stress by promoting replication fork progression. PMID: 25347734
- The results show that several leukemia-associated fusion genes cause an increase in DEK protein expression. PMID: 25524609
- DEK expression during normal hematopoiesis revealed a relationship with specific cell types, suggesting a distinct function during myeloid differentiation. PMID: 25128083
- These data suggest DEK overexpression as a critical event for the emergence of an aggressive phenotype in colorectal cancer. PMID: 25515240
- Studies demonstrate that DEK overexpression, due in part to Ron receptor activation, drives breast cancer progression through the induction of Wnt/beta-catenin signaling. PMID: 24954505
- DEK plays a significant role in the progression of small cell lung cancer. PMID: 25197373
- Results highlight a novel function of DEK as a guardian of chromatin by restricting widespread histone access, and specifically by modulating differential H3.3 loading in specific chromatin areas. PMID: 25049225
- t(6;9)/DEK-NUP214 represents a unique subtype of acute myeloid leukemia with a high risk of relapse. PMID: 24441146
- Data demonstrate that DEK stimulates head and neck squamous cell carcinoma (HNSCC) cellular growth and identify DeltaNp63 as a novel DEK effector. PMID: 24608431
- DEK is an hTERT repressor shared by various leukemia subtypes that may be involved in the deregulation of numerous genes associated with leukemogenesis. PMID: 24563617
- A high level of DEK protein expression predicts a poor prognosis for patients with gastric cancer. PMID: 24650035
- The expression of the fusion gene DEK-NUP214 leads to increased cellular proliferation. This has been shown to be dependent on the upregulation of the signal transduction protein mTOR with subsequent effects on protein synthesis and glucose metabolism. PMID: 24073922
- DEK plays a crucial role in the progression of colorectal cancers and is an independent poor prognostic factor for these cancers. PMID: 23902796
- DEK depletion inhibited cellular migration in lung cancer cell lines, possibly through inactivation of the RhoA/ROCK/MLC signal transduction pathway. PMID: 23571382
- Full-length DEK secreted by one cell can be taken up by another cell, move to the nucleus, and function in heterochromatin biology and DNA repair, potentially uniting the intracellular and extracellular activities of DEK. PMID: 23569252
- Increased expression of DEK in chronic lymphocytic leukemia correlates with immunoglobulin heavy chain variable gene mutational status, CD38-positive and del(17p13), and DEK can therefore be considered as a potential prognostic factor. PMID: 23052131
- DEK promotes the pathogenesis of estrogen receptor-positive breast cancer. PMID: 23071688
- DEK may be involved in the pathogenesis of Merkel cell carcinoma and therefore may provide therapeutic implications for these cancers. PMID: 22765016
- DEK has potent effects on hematopoietic stem cells (HSCs), hematopoietic progenitor cells (HPCs), and hematopoiesis, information of biological and potential clinical interest. PMID: 21943234
- C/EBPalpha and DEK coordinately activate myeloid gene expression. PMID: 22474248
- DEK knockdown results in a significant growth repression of CaSki cells by inducing cell apoptosis and senescence via upregulation of p65. PMID: 22390170
- DEK overexpression appears to be associated with breast cancer progression. PMID: 22360505
- DEK is important for DNA double-strand break repair. PMID: 21653549
- DEK protein is expressed in bladder tumor tissue and voided urine of bladder cancer patients. PMID: 21663673
- DEK overexpression may be a frequent event in invasive melanomas, and further augmentation of DEK expression may be associated with the acquisition of ominous features such as deep dermal invasion and metastasis. PMID: 21316078
- Data indicate that DEK expression stimulates the growth, stem cell character, and motility of breast cancer cells, and that DEK-dependent cellular invasion occurs at least in part via beta-catenin activation. PMID: 21317931
- This study identified the oncoprotein DEK, an abundant nuclear protein with a previously enigmatic in vivo function, as a Suppressor of Variegation [Su(var)] that is crucial to global heterochromatin integrity. PMID: 21460035
- DEK can contribute directly to joint inflammation in juvenile idiopathic arthritis by generating immune complexes (ICs) through high-affinity interaction between DEK and DEK autoantibodies. PMID: 21280010
- DEK overexpression, partly through an increase in its gene dose, mediates the activity of global transcriptional regulators and is associated with tumor initiation activity and poor prognosis in high-grade neuroendocrine carcinoma of the lung. PMID: 20543864
Q&A
What are the key advantages of using recombinant monoclonal antibodies over traditional animal-derived antibodies in academic research?
Recombinant monoclonal antibodies (rMAbs) offer reproducibility, cost efficiency, and ethical benefits compared to conventional animal-derived antibodies. By leveraging primary sequence data, researchers can generate standardized reagents with consistent epitope specificity, eliminating batch-to-batch variability common in commercial antibodies . Additionally, HEK293 transient gene expression (TGE) systems enable rapid production (weeks vs. months for stable cell lines) at lower costs, making large-scale antibody generation feasible for academic labs .
What is the standard workflow for producing recombinant monoclonal antibodies from primary sequences?
The process involves:
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Sequence optimization: Designing heavy (HC) and light chain (LC) geneblocks with signal peptides for secretion.
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Vector construction: Cloning sequences into mammalian expression vectors (e.g., pcDNA3.4).
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Transient transfection: Co-transfecting HEK293 cells with HC:LC ratios (e.g., 1:2 to 2:3) and polyethyleneimine (PEI) as a transfection reagent .
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Purification: Using Protein A affinity chromatography for high-purity IgG .
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Validation: Confirming binding via ELISA, Western blot, or immunofluorescence .
How do you validate the specificity of a DEK-targeting recombinant antibody?
Validation requires orthogonal approaches:
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Epitope mapping: Use peptide arrays or competition assays to confirm target binding.
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Immunoprecipitation: Verify co-purification of DEK and interacting proteins.
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Knockout controls: Test antibody binding in DEK-deficient cell lines.
How do you optimize transfection conditions for high-yield antibody production in HEK293 cells?
Critical parameters include:
For DEK rMAbs, transient expression in HEK293F cells typically yields 100–500 mg/L under optimized conditions .
What strategies address data inconsistencies when comparing recombinant vs. commercial antibodies?
Discrepancies often arise from differences in:
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Epitope accessibility: Recombinant antibodies may recognize linear vs. conformational epitopes.
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Post-translational modifications: HEK293-expressed antibodies may lack glycosylation patterns of in vivo-produced counterparts.
Solutions:
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Parallel testing: Use both antibody formats in side-by-side experiments.
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Statistical analysis: Compare EC₅₀ values in binding assays to quantify differences.
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Epitope mapping: Confirm target recognition via peptide competition .
How can recombinant antibodies be diversified for specialized applications?
Diversification strategies include:
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Species switching: Replacing constant regions (e.g., humanizing mouse antibodies) to enable multi-color imaging .
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Fragment engineering:
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scFv: Single-chain variable fragments for intracellular targeting.
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Fab: Antigen-binding fragments for minimal steric hindrance.
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Bispecific designs: Combining DEK-binding regions with secondary antigens for dual targeting .
Why might a recombinant antibody fail to bind DEK in Western blot but succeed in ELISA?
This discrepancy often stems from:
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Denaturation effects: Western blot requires SDS-PAGE, which may disrupt conformational epitopes.
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Assay sensitivity: ELISA detects linear epitopes more effectively than denatured proteins.
Resolution: Use orthogonal methods like native PAGE or live-cell imaging to confirm binding .
How do you resolve low expression yields during antibody production?
Common causes and solutions:
