REL (Ab-503) Antibody
Description
Target Biology
The REL gene encodes c-Rel, a transcription factor critical for regulating immune response genes. It forms part of the NF-κB signaling pathway, which is implicated in:
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Oncogenesis: Overexpression linked to lymphoid malignancies .
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Cellular Localization: Resides in the cytoplasm under basal conditions and translocates to the nucleus upon activation .
Key Findings Using REL (Ab-503) Antibody
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Subcellular Localization: Identified c-Rel in both nuclear and cytoplasmic compartments in human cancer cell lines using IF .
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Protein-Protein Interactions: Demonstrated complex formation between c-Rel and NF-κB subunits (e.g., p50) via co-immunoprecipitation (IP) .
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Disease Associations: Detected elevated c-Rel levels in autoimmune disorders and hematologic cancers via IHC and WB .
Validation Data
Technical Considerations
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Cross-Reactivity: Predicted reactivity with mouse c-Rel (sequence homology >95%) .
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Controls: Recommended positive controls include PMA-treated Jurkat cells .
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Limitations: Not suitable for detecting phosphorylated c-Rel (use phospho-specific antibodies for Ser503 modifications) .
References
Product Specs
Target Background
- Research suggests a possible association between cow's milk protein allergy and the cRel G+7883T polymorphism. (PMID: 29336650)
- TAB1 was identified as a functional target of miR-134, and its expression was increased by the transcription factors NF-κB1, c-Rel, and ELK1 via miR-134. (PMID: 28206956)
- NOD2 upregulates TLR2-mediated IL-23p19 expression by increasing c-Rel activation in Paneth cell-like cells. (PMID: 27563808)
- Studies demonstrate that siRNA-mediated inhibition of c-Rel expression reduces cord blood-derived B-, T-, and NK cell differentiation and expansion. (PMID: 28090796)
- Findings highlight the significance of c-REL signaling in a cellular model of cervical cancer, particularly in terms of proliferation and resistance to chemotherapeutic agents. (PMID: 28767691)
- Genetic association studies in Indian populations suggest that common polymorphisms (SNPs) in the CHGA promoter are associated with cardiometabolic disorders. c-Rel plays a role in activating CHGA promoter haplotype 2 (variant T alleles at -1018 and -57 bp) under both basal and pathophysiological conditions. (CHGA = chromogranin A; c-Rel = c-Rel proto-oncogene protein) (PMID: 28667172)
- Research indicates that c-Rel might play a role in promoting the invasion of choriocarcinoma cells through PI3K/AKT signaling. (PMID: 28259870)
- miR-574 and REL interfere with apoptosis in prostate cancer stem cells. (PMID: 27779701)
- Observations indicate that induced expression of miR-15b is modulated by c-Rel and CREB in response to JEV infection. (PMID: 26931521)
- Gene expression levels of Rel were deregulated in 49 B-cell chronic lymphocytic leukemia, 8 B-cell non-Hodgkin's lymphoma, 3 acute myeloid leukemia, 3 chronic myeloid leukemia, 2 hairy cell leukemia, 2 myelodysplastic syndrome, and 2 T-cell large granular lymphocytic leukemia patients in the post-Chernobyl period. (PMID: 25912249)
- The REL rs842647 polymorphism may be a susceptibility factor for Behcet's Disease pathogenesis and skin lesions. (PMID: 26784953)
- Analysis of c-Rel nuclear expression, REL amplification, and crosstalk between c-Rel and the p53 pathway reveals prognostic roles in diffuse large B-cell lymphoma. (PMID: 26324762)
- Overexpression of nuclear NF-κB1 and c-Rel are strong risk factors associated with chemoresistance and poor prognosis in serous epithelial ovarian cancer. (PMID: 26683819)
- Our research supports a role for c-Rel in processes crucial for keratinocyte integrity and malignant transformation, such as adhesion and migration. (PMID: 25842167)
- c-Rel is a critical mediator of NF-κB-dependent TRAIL resistance in pancreatic cancer cells. (PMID: 25299780)
- c-Rel regulates Ezh2 expression in lymphocytes and malignant lymphoid cells within a novel transcriptional network. (PMID: 25266721)
- The REL SNP rs9309331 homozygous minor allele was associated with higher LDL levels in rheumatoid arthritis. (PMID: 24489016)
- Data indicate that the NF-κB subunit c-Rel is modified and activated by O-GlcNAcylation. (PMID: 23982206)
- Our studies suggest that c-Rel is a key regulator of cell fate decisions in keratinocytes, including cell growth and death, and may play a role in epidermal carcinogenesis. (PMID: 23892589)
- Findings confirm the association of early-onset psoriasis with REL (rs13031237). (PMID: 23106574)
- An increase in the activity of the NF-κB subunit c-Rel provides protection against cell death in human islets. (PMID: 19706790)
- Activation of NF-κB p65 and c-Rel may be considered an important regulator of hypersplenism and liver cirrhosis. (PMID: 23195252)
- REL polymorphisms lack association with rheumatoid arthritis in the Tunisian population. (PMID: 22459418)
- UCP4 is a target effector gene of the NF-κB c-Rel prosurvival pathway to mitigate the effects of oxidative stress. (PMID: 22580300)
- Kidney allografts from clinical operational tolerance patients exhibit significant cellular infiltrates but a distinct expression of proteins involved in the NFkappaB1/c-rel pathway. (PMID: 22955189)
- c-Rel, as a member of the Rel/NF-κB family, is associated with psoriatic arthritis. (PMID: 22170493)
- IRF-4 was shown to enhance c-Rel-dependent binding and activation of the interleukin-4 (IL-4) promoter region. IL-2 production was also enhanced by exogenously expressed IRF-4 and c-Rel. (PMID: 21890374)
- Nuclear factor kappaB subunits RelB and cRel negatively regulate Toll-like receptor 3-mediated β-interferon production through induction of transcriptional repressor protein YY1. (PMID: 22065573)
- Levels of c-Rel directly modulated expression of caspase-4, as well as other endoplasmic reticulum stress genes. (PMID: 21984918)
- Through a novel, reversible dynamic mechanism, TNF-α-induced c-REL/ΔNp63α interactions inactivate tumor suppressor TAp73 function, promoting TNF-α resistance and cell survival in cancers with mtTP53. (PMID: 21933882)
- TAK1-c-Rel and IRF4 pathways play distinct roles in maintaining the IL-9-producing Th17 phenotype of HTLV-1-transformed cells. (PMID: 21498517)
- Three ulcerative colitis susceptibility loci are associated with primary sclerosing cholangitis, indicating a role for IL2, REL, and CARD9. (PMID: 21425313)
- Data show that Foxp3 directly or as part of a multimeric complex engages with the NF-κB component c-Rel. (PMID: 21490927)
- IL-23 induction by β-glucans is due to activation of c-Rel associated with Ser-10-histone H3 phosphorylation in the il23a promoter mediated by MAPK and SAPK or PKA, and inhibition of il12a transcription. (PMID: 21402701)
- Thus, dectin-1 and dectin-2 control adaptive T(H)-17 immunity to fungi through Malt1-dependent activation of c-Rel. (PMID: 21283787)
- The described effect of REL rs13031237 on the predisposition for rheumatoid arthritis was re-evaluated in a large case-control data set of 23,711 individuals and showed a modest increase in rheumatoid arthritis risk. (PMID: 20876593)
- Three new susceptibility loci were identified at 2p16.1 (rs1432295, REL), 8q24.21 (rs2019960, PVT1), and 10p14 (rs501764, GATA3). (PMID: 21037568)
- CXCR2 signaling is crucial in transgenic mice with C-rel-deficient/NFkappaB1-deficient/heterozygous Rela+/- neutrophilia, causing spontaneous inflammation. (PMID: 20519647)
- The REL locus is associated with rheumatoid arthritis susceptibility in the UK population. (PMID: 19945995)
- This study aimed to (i) characterize the prevalence of REL, BCL11A, and MYCN gains in a consecutive CLL series at the time of diagnosis; (ii) define the prognostic relevance of REL, BCL11A, and MYCN gains in CLL. (PMID: 20575024)
- c-Rel, but not nuclear factor-κB1 (NFKB1), is required for the development of transgenic regulatory T cell progenitors. (PMID: 20228198)
- The kinetics of NFkappaB subunit activation are partly responsible for the observed pattern of acute inflammation in the adenoviral-infected cornea. (PMID: 20038977)
- REL, rather than BCL11A, may be the target of the 2p13 alterations in classical Hodgkin Lymphoma. (PMID: 11830502)
- REL plays a significant pathological role in Hodgkin's lymphoma. (PMID: 12478664)
- The correlation of structural aberrations of the REL locus and nuclear c-Rel accumulation in Reed-Sternberg cells qualifies REL as a target gene of the frequent gains in 2p in cHL. REL aberrations contribute to constitutive NF-κB/Rel activation in cHL. (PMID: 12511414)
- Calmodulin binds c-Rel and RelA after their release from IκB and can inhibit nuclear import of c-Rel while allowing RelA to translocate to the nucleus and act on its target genes. (PMID: 12556500)
- v-REL plays a role in NF-κB-regulated cell death. (PMID: 12588973)
- Deletion of either C-terminal transactivation subdomain enhances the in vitro transforming activity of REL in chicken spleen cells. (PMID: 14534540)
- REL amplification may not be causative in diffuse large B-cell lymphoma. (PMID: 14615382)
- Rel/NF-κB factors could participate in the occurrence of senescence by generating oxidative stress. (PMID: 14744759)
Q&A
What experimental applications are most suitable for REL (Ab-503) Antibody in cellular studies?
REL (Ab-503) Antibody is primarily utilized for detecting phosphorylated c-Rel at serine 503 in human cells via immunofluorescence (IF). Its specificity for the phosphoepitope (T-S-S(p)-D-S) makes it ideal for:
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Mapping spatiotemporal activation of NF-κB signaling in response to cytokines (e.g., TNF-α, IL-1β) or cellular stress .
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Investigating c-Rel nuclear translocation dynamics in immune cells (e.g., lymphocytes, macrophages) under inflammatory conditions .
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Validating CRISPR/Cas9-mediated phosphorylation site mutations in REL gene-edited cell lines .
Table 1: Key Applications and Protocols
How can researchers rigorously validate the specificity of REL (Ab-503) Antibody?
Specificity validation requires a multi-modal approach:
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Peptide Blocking Assays: Pre-incubate the antibody with its immunogen peptide (T-S-S(p)-D-S). A >90% reduction in signal confirms epitope specificity .
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Knockout Controls: Use REL−/− cell lines (e.g., CRISPR-edited HEK293) to confirm absence of non-specific binding .
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Cross-Reactivity Screening: Test against recombinant proteins of other Rel/NF-κB family members (e.g., RelA, RelB) to rule out off-target interactions .
What methodological considerations are critical for optimizing REL (Ab-503) in time-course experiments?
Phosphorylation at Ser503 is transient, peaking 15–30 minutes post-stimulation (e.g., LPS treatment) and declining by 2 hours . To capture this dynamic:
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Fixation Timing: Terminate experiments at 15-minute intervals post-induction.
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Inhibition Cocktails: Include phosphatase inhibitors (e.g., sodium orthovanadate) in lysis buffers to preserve phospho-signals .
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Multiplex Imaging: Pair with antibodies against IκBα degradation to correlate c-Rel activation with upstream pathway events .
How should researchers resolve contradictory staining patterns in heterogeneous cell populations?
Contradictory signals often arise from:
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Cell Cycle-Dependent Phosphorylation: c-Rel phosphorylation varies across phases; synchronize cells using double thymidine block .
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Subpopulation Analysis: Combine IF with flow cytometry using lineage markers (e.g., CD4+ T cells vs. macrophages) .
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Quantitative Thresholds: Establish fluorescence intensity cutoffs via histogram analysis of isotype controls .
Case Study: A 2024 trial reported false-positive nuclear staining in senescent fibroblasts, resolved by adding 0.1% Triton-X for improved cytoplasmic washout .
What advanced techniques complement REL (Ab-503) for mechanistic studies?
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Proximity Ligation Assay (PLA): Visualize c-Rel interaction partners (e.g., p50) within 40 nm resolution .
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CUT&Tag Sequencing: Map genome-wide c-Rel binding sites post-phosphorylation using Cleavage Under Targets and Tagmentation .
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Phos-Tag™ Electrophoresis: Resolve phosphorylated vs. total c-Rel in stimulated vs. naive cells .
Table 2: Integrated Workflow for NF-κB Pathway Analysis
| Step | Technique | Key Readout |
|---|---|---|
| 1. Stimulation | TNF-α (10 ng/mL, 15 min) | IκBα degradation |
| 2. Localization | REL (Ab-503) IF | Nuclear translocation |
| 3. DNA Binding | ChIP-seq | Target gene occupancy |
What are the implications of Ser503 phosphorylation in disease models?
Ser503 modification regulates c-Rel’s transcriptional activity in:
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Autoimmunity: Elevated phosphorylation in rheumatoid arthritis synovium correlates with IL-17A overexpression .
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Oncology: Constitutive phosphorylation in diffuse large B-cell lymphoma enhances anti-apoptotic BCL-2 expression .
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Validation Strategy: Compare phospho-c-Rel levels in patient-derived xenografts vs. healthy controls using quantitative IF analysis .
How does batch-to-batch variability impact reproducibility, and how can it be mitigated?
While polyclonal antibodies inherently exhibit variability, REL (Ab-503) batches show <15% signal variance when:
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Normalization: Pre-titer each lot against a reference sample (e.g., LPS-treated THP-1 cells) .
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Cross-Validation: Confirm key findings with orthogonal methods (e.g., phospho-specific Western blot) .
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Long-Term Storage: Aliquot in 50% glycerol, avoiding >3 freeze-thaw cycles to preserve affinity .
What computational tools enhance analysis of REL (Ab-503) datasets?
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QuPath: Quantify nuclear vs. cytoplasmic staining intensity in whole-slide images .
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CellProfiler: Classify phosphorylation states using machine learning pipelines trained on Ser503 mutants .
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STRING-DB: Predict c-Rel interaction networks perturbed by Ser503 phosphorylation .
How does REL (Ab-503) compare to commercial alternatives for phospho-c-Rel detection?
Table 3: Performance Comparison
| Antibody | Vendor | Sensitivity (IF) | Cross-Reactivity |
|---|---|---|---|
| REL (Ab-503) | Abeomics | 1:200 | RelA (weak) |
| Phospho-c-Rel (S503) | Cell Signaling | 1:500 [N/A] | None reported |
| c-Rel pS503 | Sigma-Aldrich | 1:100 | RelB (moderate) |
Note: Independent validation using siRNA knockdown is recommended for all commercial antibodies .
