Customize lmo3 Antibody

Description

LMO3 Protein Overview

Structure:

LMO3 belongs to the LIM-only (LMO) protein family, characterized by two LIM zinc-binding domains that mediate protein-protein interactions, particularly with transcription factors like HEN2 and LATS1 .
Functions as a transcriptional co-regulator, influencing pathways such as Hippo signaling, epithelial-mesenchymal transition (EMT), and immune response .

Expression Patterns:

Predominantly expressed in neuroendocrine tissues but downregulated in multiple cancers, including pancreatic ductal adenocarcinoma (PDAC) and prostate cancer (PCa) .
Reduced LMO3 correlates with advanced tumor stage, higher pathological grade, and poor survival in PDAC .

Research Applications of LMO3 Antibodies

LMO3 antibodies enable:

Immunohistochemistry (IHC): Detects cytoplasmic LMO3 in acinar/endocrine cells and tumor tissues (Supplementary Figure 1A-B) .
Western Blotting: Validates LMO3 overexpression in PDAC cell lines (e.g., Panc 10.05, SU.86.86) .
Transcriptome-Metabolome Integration: Identifies LMO3-associated metabolic reprogramming (e.g., glycerol 3-phosphate accumulation) .

Table 1: LMO3 Expression and Clinical Outcomes

Cancer Type	LMO3 Role	Clinical Correlation	Source
Pancreatic Cancer	Suppresses basal-like/squamous subtype	↑ LMO3 → ↑ survival (HR = 0.40)
Prostate Cancer	Downregulated in tumors	↓ LMO3 → ↑ immune evasion

Table 2: Functional Pathways Regulated by LMO3

Pathway	Effect of LMO3 Upregulation	Experimental Model
Cellular proliferation	Inhibited (CCK-8/WST-8 assays)	PDAC cell lines
Migration/Invasion	Reduced (Transwell assays)	SU.86.86 cells
Immune infiltration	↑ Mast cells, NK cells, Th1 cells	PCa (TIMER/ssGSEA)

Mechanistic Insights

Metabolic Reprogramming:
- LMO3 upregulation enhances lipogenesis and glycerol 3-phosphate (G3P) synthesis via GPD1, suppressing amino acid metabolism in PDAC .
- High G3P levels correlate with improved survival (HR = 0.39, P < 0.05) .
Immune Modulation:
- In PCa, LMO3 downregulation associates with elevated PD-1, PD-L1, and CTLA-4 expression, promoting immune evasion .
- LMO3-high tumors exhibit stromal/immune cell infiltration (↑ macrophages, dendritic cells) .

Therapeutic and Diagnostic Implications

Biomarker Potential:
- Low LMO3 predicts aggressive PDAC subtypes and poor response to methotrexate/vinblastine in PCa .
- High LMO3 correlates with sensitivity to anti-PD-1 immunotherapy (IPS score ↑) .
Targeted Therapy:
- Restoring LMO3 expression inhibits PDAC proliferation and basal-like differentiation .
- Combinatorial approaches with immune checkpoint inhibitors (e.g., anti-CTLA-4) may benefit LMO3-low PCa .

Limitations and Future Directions

Current studies rely on mRNA data; protein-level validation is needed .
Mechanistic links between LMO3 and metabolic/immune pathways require in vivo confirmation .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

lmo3 antibody; zgc:110149 antibody; LIM domain only protein 3 antibody; LMO-3 antibody

Target Names

lmo3

Uniprot No.

Q503U0

Q&A

What criteria should guide LMO3 antibody selection for immunohistochemistry (IHC) versus Western blot (WB)?

Antibody performance depends on epitope accessibility, host species compatibility, and validation evidence. For IHC, prioritize antibodies validated for formalin-fixed paraffin-embedded (FFPE) tissues with cytoplasmic/nuclear localization data . The 2H2 clone (mouse monoclonal IgG2a) demonstrates specificity for AA 91-146 in human FFPE sections , while polyclonal antibodies like PACO21903 (rabbit) show broader species reactivity (human/mouse/rat) . For WB, confirm antibody recognition of denatured epitopes: the 4A8 clone (IgG2b) detects recombinant LMO3 at 16.6 kDa , whereas goat polyclonal antibodies (abx617313) target C-terminal domains with 1:128,000 dilution efficacy in P-ELISA .

Table 1: Antibody validation parameters across platforms

Clone	Host	Applications	Epitope Region	Validation Evidence
2H2	Mouse	IF, IHC, WB	AA 91-146	Recombinant protein controls
1A8	Mouse	IP, IF, ELISA	Full-length	Neuroblastoma cell lysates
abx617313	Goat	FCM, IF/ICC	C-terminal	Peptide blocking assays

How should researchers validate LMO3 antibody specificity in novel experimental systems?

Implement a three-tier validation framework:

Genetic controls: Compare signal intensity in LMO3-knockout vs. wild-type cell lines (e.g., CRISPR-edited Panc 10.05 PDAC cells) .
Orthogonal verification: Correlate IHC results with RNAscope® in situ hybridization or qRT-PCR . In PCa studies, LMO3 mRNA-protein expression correlation reaches R = 0.78 (P < 0.001) .
Epitope mapping: Use truncated recombinant proteins (e.g., GST-tagged NP_061110 fragments) in dot blot assays . The 2H2 clone shows 92% sequence coverage across LMO3 isoforms .

How can conflicting reports about LMO3's oncogenic versus tumor-suppressive roles be resolved?

Context-dependent LMO3 functions require careful experimental design:

Step 1: Subtype stratification
In PDAC, LMO3 suppresses basal-like/squamous subtypes (HR = 0.40, P < 0.05) but shows neutral effects in classical subtypes . Always stratify analyses using consensus molecular subtyping (e.g., Moffitt classification) .

Step 2: Metabolic contextualization
LMO3-high PDAC tumors exhibit:

2.1-fold increase in glycerol-3-phosphate (G3P) (P = 0.007)
58% reduction in glutamine uptake (P = 0.03)
Profile metabolomes using LC-MS alongside transcriptomics to resolve phenotypic contradictions.

Step 3: Pathway enrichment analysis
In PCa, LMO3 correlates with:

Immune infiltration (CD8+ T cells: R = 0.31, P = 0.002)
Stromal activation (TGF-β pathway: P = 3.2e-5)
Use ssGSEA to separate immune-mediated versus cell-autonomous effects .

What multi-omics strategies optimize LMO3 functional studies in PDAC progression models?

Integrate these approaches for mechanistic insights:

A. Transcriptome-metabolome coupling
In LMO3-overexpressing PDAC cells:

RNA-seq identifies 127 downregulated basal-like genes (FDR < 0.05)
Metabolomics reveals 11.3 μM G3P accumulation (P = 0.004) via GPD1 upregulation
ChIP-seq maps LMO3 binding to GPD1 promoter (3.8-fold enrichment)

B. Spatiotemporal resolution
Employ multiplex IHC panels with antibodies against:

LMO3 (clone 2H2 )
GPD1 (validated in )
Ki-67 (proliferation marker)
Quantify spatial co-localization using HALO® image analysis.

How should researchers address technical variability in LMO3 quantification across antibody batches?

Implement a standardized validation protocol:

Parameter	QC Criteria	Corrective Action
Lot-to-lot consistency	<15% CV in ELISA titers	Parallel testing with NIST-like reference material
Epitope integrity	95% sequence coverage by MS	Epitope mapping via HDX-MS
Cross-reactivity	≤5% signal in KO models	Pre-adsorption with blocking peptides

For longitudinal studies, aliquot master stocks at -80°C with 0.02% sodium azide .

What statistical approaches reconcile LMO3 expression discrepancies between TCGA and GEO datasets?

Apply meta-analysis frameworks:

A. Effect size harmonization
For PCa data:

Dataset	N	LMO3 Δ (Tumor vs. Normal)	95% CI
TCGA	499	-1.8-fold	[-2.1, -1.5]
GSE30994	64	-1.2-fold	[-1.5, -0.9]
GSE70769	112	-2.3-fold	[-2.7, -1.9]

Use random-effects models (DerSimonian-Laird) to calculate pooled estimate: -1.7-fold (95% CI: -2.0 to -1.4) .

B. Batch effect correction
Apply ComBat algorithm to microarray/RNA-seq data, preserving biological variance while removing platform-specific biases .

Can LMO3 antibody-based assays stratify PDAC patients for targeted therapy?

Emerging evidence supports LMO3 as a theranostic biomarker:

Predictive value

LMO3 Status	Median OS (Months)	HR (95% CI)
High	28.4	0.39 (0.22-0.67)
Low	14.7	Reference

Data from NCI-UMD-German cohort (n=152) . Validate with orthogonal methods:

Circulating tumor DNA methylation at LMO3 locus (ddPCR sensitivity: 0.01% allele frequency)
Multiplexed ion beam imaging (MIBI) for protein co-expression networks

What mechanistic studies elucidate LMO3's role in transcriptional regulation?

Employ these experimental pipelines:

Co-factor identification

Co-IP with LMO3 antibody (1A8 clone )
Mass spectrometry identifies binding partners (e.g., LDB1, NKX2-5)
CRISPRi screening to map genetic interactors

B. 3D chromatin architecture
Combine CUT&RUN (using LMO3 antibody) with Hi-C to resolve looping interactions at:

GPD1 locus (chr12:16.5 Mb)
Basal-like signature genes (KRT5, DSG3)

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lmo3 Antibody