DEFB1 Recombinant Monoclonal Antibody
Description
Introduction to DEFB1 Recombinant Monoclonal Antibody
DEFB1 recombinant monoclonal antibodies are laboratory-generated proteins that specifically bind to human β-defensin-1, a peptide encoded by the DEFB1 gene. These antibodies enable precise detection of DEFB1 in research settings, aiding studies on its antimicrobial functions, regulatory pathways, and roles in diseases such as viral infections and cancer .
Development and Production
Key characteristics of DEFB1 recombinant monoclonal antibodies include:
These antibodies undergo rigorous purification processes, including affinity chromatography, to ensure high specificity .
Validation and Specificity
DEFB1 recombinant monoclonal antibodies are validated across multiple platforms:
Table 1: Validation Data
Specificity is confirmed using DEFB1-knockout controls and siRNA silencing .
Applications in Research
DEFB1 recombinant monoclonal antibodies are used to:
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Study Viral Infections: Quantify DEFB1’s suppression of influenza A replication in bronchial epithelial cells via STAT3 modulation .
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Investigate Cancer Pathways: Analyze reduced DEFB1 expression in colorectal cancer organoids and its link to tumor suppression .
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Explore Urinary Tract Immunity: Assess DEFB1’s role in combating E. coli infections in bladder and kidney tissues .
Research Findings Using DEFB1 Recombinant Monoclonal Antibody
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Influenza Regulation: Overexpression of DEFB1 in bronchial cells reduced influenza A(H1N1) matrix gene copies by 99%, demonstrating antiviral activity .
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Cancer Signaling: Loss of DEFB1 in colon cancer correlates with disrupted NF-κB and STAT3 pathways, implicating it in tumor suppression .
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Bacterial Defense: DEFB1 knockout mice showed altered E. coli clearance rates in urinary tract infections, highlighting its context-dependent antimicrobial role .
Product Specs
The DEFB1 recombinant monoclonal antibody is produced through in vitro expression systems. The DNA sequences encoding DEFB1 antibodies, sourced from immunoreactive rabbits, are cloned and inserted into plasmid vectors. These recombinant plasmid vectors are subsequently transfected into host cells for antibody expression. The immunogen used in this process is a synthesized peptide derived from the human DEFB1 protein. Following expression, the DEFB1 recombinant monoclonal antibody undergoes affinity-chromatography purification and is rigorously tested for functionality in ELISA and FC applications. These tests demonstrate reactivity with the human DEFB1 protein, confirming its ability to bind and interact with the target antigen.
DEFB1, a human beta-defensin, is an antimicrobial peptide that plays a crucial role in the innate immune response. It provides defense against a wide range of microbial pathogens, including bacteria, fungi, and viruses. Its presence at mucosal surfaces, particularly those exposed to the external environment, underscores its essential role in safeguarding the body against infections.
Target Background
DEFB1 exhibits bactericidal activity. It may also act as a ligand for the C-C chemokine receptor CCR6. This interaction positively regulates sperm motility and bactericidal activity in a CCR6-dependent manner. DEFB1 binds to CCR6 and triggers Ca2+ mobilization within sperm, a process crucial for sperm motility.
- An association between the allelic distribution of the rs12355840 polymorphism in miRNA202 and caries experience was observed in the Brazilian Manaus group. In the Ribeirao Preto group, the allelic and genotypic distributions of the rs11362 polymorphism in DEFB1 were associated with caries experience. PMID: 29727722
- These studies identified SNPs that served as cis-eQTLs for DEFB1 and AHR and were associated with variations in plasma KYN concentrations, which were linked to the severity of major depressive disorder symptoms. PMID: 29317604
- A significant connection was observed between DEFB1 and TCF7L2 gene polymorphisms and nephrolithiasis. PMID: 29959006
- These findings identified critical promoter regions of DEFB1 that are epigenetically influenced by lactobacilli in vaginal keratinocytes. PMID: 28508168
- Endometrial tissues and epithelial cells constitutively secreted both BD-1 and -2, primarily at the apical compartment. Both genistein and daidzein induced BDs secretion, reaching a peak at 5-15 minutes. The apical secretion of BDs coincided with increased Cl- secretion induced by genistein. PMID: 29905453
- The polymorphisms in DEFB1 were not associated with salivary hBD levels or caries experience (p > 0.05) in children. PMID: 28343232
- These findings suggest a potential role for hBD-1 in the pathogenesis of psoriasis. PMID: 28067065
- Significant associations were found between periodontitis and g. -20G> A (rs11362) and g. -44C> G (rs1800972) SNPs in the DEFB1 gene, as well as p.Ala29Thr (rs1126477) and p.Lys47Arg (rs1126478) SNPs in the LTF gene. PMID: 28485077
- hBD-1 potentiates the induction of in vitro osteoclastogenesis by RANKL via enhanced phosphorylation of the p44/42 MAPKs. PMID: 28709835
- These findings suggest a potential involvement of hBD-1 mediated innate immunity in modulating susceptibility to the development of adeno-tonsillar hypertrophy. PMID: 29501294
- Two DEFB1 SNPs were significantly associated with the DMFT (decayed, missing, filled teeth) index. PMID: 27846636
- Multivariate logistic regression analysis indicated that hBD-1 basal cell loss (>/=20% of prostatic glands in total cores) is an independent factor predicting prostatic adenocarcinoma (odds ratio: 4.739, confidence interval: 1.093-20.554, p = 0.038). Loss of hBD-1 basal cells serves as a valuable indicator to identify patients at extremely high risk with initially negative biopsy results. PMID: 28885732
- The data highlight the differential regulation of colonic epithelial beta-defensin expression and secretion by bile acids. This study discusses the implications for intestinal health and disease. PMID: 28487283
- This research provides valuable insights into the in vivo kinetics of HBD expression, the mechanism of HBD1 induction by HIV-1, and the role of HBDs in the early innate response to HIV-1 during acute infection. PMID: 28253319
- CpG methylation frequencies in the DEFB1 low CpG-content promoter were significantly higher in prostate malignant tissues compared to adjacent benign tissues. PMID: 27835705
- DEFB1 genotypes were correlated with the salivary concentration of hBD-1. PMID: 27770642
- Coding missense and nonsense single nucleotide polymorphisms result in reduced antibacterial activity of HBD1 against Escherichia coli. PMID: 27160989
- A significant difference was observed between the levels of staining of TLR-2, TLR-4, and hBD-1 in different lesions from the epidermis, inflammation region, dermis, and skin appendages (p < 0.05) in patients with acne vulgaris. PMID: 26695933
- Expression of hBD-1, hBD-2, hBD-3, and hBD-4 was examined in healthy and chronic periodontitis gingiva. PMID: 26874342
- The -44 CC genotype of the beta-defensin-1 gene (DEFB1 rs1800972) may be associated with susceptibility to chronic periodontitis in Japanese individuals. PMID: 24276427
- No clear association was found between DEFB1 polymorphisms and tuberculosis. PMID: 26991287
- While DEFB1 polymorphisms alone may not influence pathology susceptibility, they could potentially contribute, alongside other factors involved in the genetic control of the innate immune system, to the predisposition towards recurrent tonsillitis. PMID: 26968045
- Beta-defensin-1 production is dysregulated in the epithelium of patients with chronic obstructive pulmonary disease. PMID: 26568662
- Dental plaque amount, lactobacilli count, age, saliva buffer capacity, and DEFB1 gene polymorphism accounted for a total of 87.8% of variations in DMFT scores. PMID: 26377569
- This meta-analysis provides evidence that DEFB1 genetic polymorphisms rs11362G>A, rs1800972C>G, and rs1799946G>A are significant contributing factors to the development of digestive diseases. PMID: 26232989
- The association of infectious pathology of the upper respiratory tract and the development of periodontitis diseases with markers in DEFB1 (-44G/C) and TLR2 Arg753Gln and Arg677Trp genes was determined. PMID: 27029120
- Genotypes and alleles of the DEFB1 gene identified in long-living individuals can be considered as factors that increase the probability of longevity and a favorable course of age-related diseases. PMID: 26028230
- There is a significant link between innate immunity deregulation, through disruption of cationic peptides (hBDs), and the potential development of colon cancer. PMID: 26038828
- A possible association between DEFB1 polymorphisms and in vivo salivary levels of hBD-1 was explored in an Italian population of 40 healthy individuals. The study found a genotype-related differential expression with two of these polymorphisms: -52G > A and -44C > G. PMID: 25939140
- RNA interference-induced Caspase-12 silencing was found to increase NOD1, hBD1, and hBD2 expression. PMID: 25503380
- A-dependent DEFB1 upregulation below 20 mM predicts in vitro antimicrobial activity, as well as glucose- and AA-dependent CAMP and IFNG upregulation. PMID: 25815330
- Plant-made recombinant beta-defensins (phBD-1 and phBD-2) are promising antimicrobial substances with the potential to become additional tools against salmonellosis, particularly when used in combination. PMID: 25417183
- The DEFB1 c.-44C>G polymorphism of the GG protective genotype was significantly less frequent among multiple sclerosis patients compared to control groups. PMID: 26434201
- Beta defensin 1 gene -20G/A, -44C/G, and -52G/A single nucleotide polymorphisms were not associated with chronic tonsillitis. PMID: 25683590
- These results suggest the involvement of DEFB1 polymorphisms, particularly that of the c.5*G>A SNP, in the susceptibility of women from Northeast Brazil to HPV infection. PMID: 25410996
- Functional polymorphisms in IL1B and DEFB1 influence susceptibility to mold infection in solid-organ transplant recipients. PMID: 25398456
- DEFB1 minor haplotypes were also associated with an increased risk of developing Atypical Squamous Cells of Undetermined Significance lesions, being significantly more frequent in HPV negative women with lesions than without lesions. PMID: 24435641
- This finding explains a common defect in male infertility associated with both asthenozoospermia and leukocytospermia, indicating a dual role of DEFB1 in defending male fertility. PMID: 25122636
- The GG genotype of the C-44G SNP in the DEFB1 gene may result in decreased defensin beta-1 production in diabetes. PMID: 25083086
- These findings associate DEFB1 5'UTR polymorphisms with HIV-1/AIDS in Mexican women for the first time. PMID: 25001249
- hBD-1 suppresses tumor migration and invasion of oral squamous cell carcinoma. PMID: 24658581
- Data indicate that CCL2 and IL-10 released from burn patient lineage-CD34+CD31- cells were shown to be responsible for their inhibitory activities on the production of beta-defensin 1 (HBD-1) by epidermal keratinocytes. PMID: 24498256
- A possible involvement of the DEFB1 gene in inflammatory bowel diseases is suggested. PMID: 24034632
- A genetic variant, rs11362, in DEFB1 influences caries susceptibility in CL/P children. These results support the hypothesis that DEFB1 expression in saliva may serve as a biomarker for future caries risk. PMID: 23964634
- This gene encodes a peptide with antimicrobial activity against S. aureus and H. influenzae. PMID: 20100591
- HBD-1 serum levels in chronic kidney disease patients are inversely related to estimated glomerular filtration rate. PMID: 24356550
- These findings suggest an interplay between hBD-1 and neuroimmunological responses in Alzheimer's disease. PMID: 24139179
- IRF5, but not TLR4, DEFEB1, or VDR, is associated with the risk of ulcerative colitis in a Han Chinese population. PMID: 23971939
- Data suggest that levels of DEFB1 and DEFB2 are highest in colostrum and decrease in human milk over the course of lactation. The abundance of DEFB1 in colostrum may provide protection against upper respiratory infection in infants up to 6 months. PMID: 24124699
- The salivary beta-defensin concentration was significantly higher in patients with oral mucosal diseases than in healthy volunteers. Furthermore, in patients, the concentration was significantly higher before treatment than after treatment. PMID: 24564045
Q&A
What is the biological function of DEFB1, and why is it significant in immunological research?
DEFB1, also known as beta-defensin 1, is a small cationic peptide that plays a crucial role in the innate immune system. It exhibits antimicrobial activity against a broad spectrum of pathogens, including bacteria, fungi, and viruses. DEFB1 is expressed in epithelial cells and serves as a first line of defense by disrupting microbial membranes through its amphipathic structure . Additionally, it has been implicated in modulating immune responses by acting as a ligand for the C-C chemokine receptor CCR6, which triggers calcium mobilization and enhances sperm motility .
In immunological research, DEFB1 is studied for its potential role in host-pathogen interactions, inflammatory diseases, and cancer. For instance, its expression levels have been correlated with susceptibility to infections and autoimmune disorders such as Crohn's disease. Understanding the mechanisms underlying DEFB1 activity can provide insights into therapeutic strategies for enhancing innate immunity or modulating inflammatory responses .
How can recombinant monoclonal antibodies against DEFB1 be utilized in experimental research?
Recombinant monoclonal antibodies against DEFB1 are invaluable tools for studying its expression, localization, and function. These antibodies are engineered to have high specificity and affinity for DEFB1, making them suitable for various applications such as Western blotting (WB), immunohistochemistry (IHC), enzyme-linked immunosorbent assays (ELISA), and flow cytometry .
For example:
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Western Blotting: Researchers can use these antibodies to detect DEFB1 protein levels in cell lysates or tissue extracts. This application is particularly useful for quantifying changes in DEFB1 expression under different experimental conditions.
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Immunohistochemistry: By staining tissue sections with anti-DEFB1 antibodies, researchers can visualize the spatial distribution of DEFB1 within specific cell types or tissue compartments.
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Flow Cytometry: This technique allows for the analysis of DEFB1 expression at the single-cell level, facilitating studies on cell-specific expression patterns or functional assays involving immune cells.
What are the key considerations when designing experiments using anti-DEFB1 recombinant monoclonal antibodies?
Experimental design involving anti-DEFB1 recombinant monoclonal antibodies requires careful attention to several factors:
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Antibody Validation: Ensure that the antibody has been validated for the intended application (e.g., WB, IHC). Check validation data provided by manufacturers or published studies to confirm specificity and sensitivity .
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Positive and Negative Controls: Include appropriate controls to validate antibody performance. For instance, use tissues or cells known to express DEFB1 as positive controls and those lacking DEFB1 expression as negative controls.
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Optimization of Conditions: Determine the optimal antibody concentration and incubation conditions for your assay. This may involve titration experiments to identify the concentration that provides the best signal-to-noise ratio.
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Blocking Steps: Use blocking agents (e.g., bovine serum albumin or non-fat milk) to minimize non-specific binding.
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Species Reactivity: Verify that the antibody is reactive with the species being studied. For example, some anti-DEFB1 antibodies are specific to human samples but may not cross-react with mouse or rat proteins .
By addressing these considerations, researchers can enhance the reliability and interpretability of their experimental results.
How does one interpret contradictory data when studying DEFB1 using recombinant monoclonal antibodies?
Contradictory data can arise from various sources when studying DEFB1. To address such inconsistencies:
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Reevaluate Antibody Specificity: Perform additional validation experiments to confirm that the antibody specifically binds to DEFB1. This may involve using blocking peptides or knockout models where DEFB1 is genetically ablated.
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Standardize Experimental Conditions: Ensure that all experiments are conducted under standardized conditions to minimize variability. Factors such as sample preparation, antibody dilution, and detection methods should be consistent across replicates.
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Consider Biological Variability: Recognize that biological systems are inherently variable. Differences in DEFB1 expression may reflect physiological or pathological states rather than experimental artifacts.
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Cross-Validate Findings: Use complementary techniques (e.g., qPCR for mRNA levels) to corroborate protein-level data obtained with recombinant monoclonal antibodies.
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Consult Literature: Compare your findings with published studies to identify potential reasons for discrepancies. Differences in experimental design or sample characteristics may explain conflicting results.
By systematically addressing these factors, researchers can resolve contradictions and gain a clearer understanding of DEFB1 biology.
What are the advantages of using recombinant monoclonal antibodies over polyclonal antibodies for studying DEFB1?
Recombinant monoclonal antibodies offer several advantages over polyclonal antibodies:
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Specificity: Monoclonal antibodies recognize a single epitope on the target protein, reducing the likelihood of cross-reactivity with other proteins.
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Reproducibility: Recombinant production ensures batch-to-batch consistency, which is critical for reproducible research.
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Customization: Recombinant technology allows for customization of antibody properties (e.g., affinity maturation or engineering of Fc regions).
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Scalability: Recombinant production enables large-scale manufacturing without relying on animal hosts.
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Validation Across Applications: Recombinant monoclonal antibodies are often validated across multiple applications (e.g., WB, IHC), providing versatility in experimental design .
These advantages make recombinant monoclonal antibodies a preferred choice for high-quality research on DEFB1.
How can researchers assess the functional role of DEFB1 in disease models using recombinant monoclonal antibodies?
To investigate the functional role of DEFB1 in disease models:
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Expression Analysis: Use anti-DEFB1 antibodies to quantify protein levels in disease versus control samples. For example, elevated DEFB1 expression may be associated with inflammatory conditions or certain cancers .
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Localization Studies: Perform IHC or immunofluorescence staining to determine whether changes in DEFB1 localization correlate with disease progression.
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Functional Assays: Conduct functional assays (e.g., bacterial killing assays) to assess whether altered DEFB1 levels impact its antimicrobial activity.
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In Vivo Studies: Use animal models genetically modified to overexpress or knock out DEFB1. Anti-DEFB1 antibodies can be used to monitor protein expression in these models.
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Pathway Analysis: Investigate downstream signaling pathways activated by DEFB1 using techniques such as phospho-specific Western blotting or reporter assays.
These approaches provide a comprehensive framework for elucidating the role of DEFB1 in health and disease.
What challenges might arise when working with anti-DEFB1 recombinant monoclonal antibodies?
Challenges associated with anti-DEFB1 recombinant monoclonal antibodies include:
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Epitope Masking: In some cases, the epitope recognized by the antibody may be masked due to post-translational modifications or protein-protein interactions.
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Non-Specific Binding: Despite high specificity, non-specific binding can occur under suboptimal assay conditions.
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Batch Variability: While recombinant production minimizes variability, differences between production batches can still occur if quality control measures are not stringent.
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Limited Cross-Reactivity: Some recombinant monoclonal antibodies may not cross-react with orthologous proteins from other species.
To mitigate these challenges, researchers should perform rigorous validation experiments and consult technical support from antibody suppliers when needed.
