DLX1 Antibody

What is DLX1 and what are its primary biological functions?

DLX1 is a homeobox transcription factor belonging to the DLX family that plays essential roles in embryonic development. It is particularly critical for the production of forebrain GABAergic interneurons during embryonic development. The DLX family of homeobox transcription factors (including DLX1, DLX2, DLX5, and DLX6) are expressed in overlapping domains at different developmental stages of cell differentiation in the subpallium and control the differentiation of GABAergic neurons. DLX1 also appears to have roles in craniofacial patterning and has been implicated in various cancer types .

What are the molecular characteristics of DLX1 protein?

DLX1 is characterized by the following molecular properties:

• Full name: distal-less homeobox 1

• Calculated molecular weight: 255 amino acids, 27 kDa

• Observed molecular weight in laboratory applications: 27 kDa

• GenBank accession number: BC036189

• Gene ID (NCBI): 1745

• UniProt ID: P56177

What sample types can be analyzed using DLX1 antibody?

Based on validated research applications, DLX1 antibody has been tested for reactivity with the following samples:

For Western Blot applications, positive detection has been confirmed in human brain tissue, A375 cells, and HeLa cells. For immunohistochemistry, positive detection has been reported in mouse brain tissue .

What are the optimal dilution parameters for DLX1 antibody in different experimental applications?

The following dilution guidelines have been established for DLX1 antibody applications:

It is important to note that these are general guidelines, and the antibody should be titrated in each specific testing system to obtain optimal results, as performance can be sample-dependent .

What antigen retrieval methods are recommended for DLX1 immunohistochemistry?

For optimal immunohistochemical detection of DLX1, the suggested antigen retrieval method is using TE buffer at pH 9.0. As an alternative approach, antigen retrieval may also be performed with citrate buffer at pH 6.0. The choice between these methods may depend on tissue type and fixation conditions .

What storage conditions maintain DLX1 antibody stability and activity?

For proper maintenance of DLX1 antibody:

• Store at -20°C

• The antibody remains stable for one year after shipment when stored properly

• Aliquoting is unnecessary for -20°C storage

• The antibody is supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.3

• Some preparations (20μL sizes) contain 0.1% BSA

How can proper validation of DLX1 antibody be ensured for experimental applications?

To ensure proper validation:

• Include positive controls such as human brain tissue, A375 cells, or HeLa cells for Western blot applications

• Use mouse brain tissue as a positive control for immunohistochemistry

• Consider knockout validation approaches, such as performing DLX2 ChIP-seq on wild-type and DLX2-/- samples (as demonstrated in the literature for related DLX proteins)

• Employ blocking peptides (the antigen used in antibody generation) as negative controls in ChIP-seq and other applications to confirm specificity

How can DLX1 antibody be utilized in chromatin immunoprecipitation (ChIP) studies?

DLX1 antibody has been successfully used in ChIP-seq applications to study genome-wide DLX binding patterns. When designing ChIP experiments:

• Ensure antibody specificity using appropriate controls, including IgG and blocking peptides specific to the DLX1 antigen

• Generate biological replicates to confirm reproducibility of binding patterns

• Consider using a tagged version of DLX1 (such as DLX1-FLAG) for enhanced specificity in pull-down experiments

• Be aware that DLX1 binding often overlaps with other DLX family members (DLX2, DLX5), with correlation of normalized ChIP-seq signals showing pairwise r² values > 0.7

• Consider analyzing different threshold levels (low, medium, and high affinities) for peak calling to identify both shared and unique binding sites

What insights can DLX1 antibody provide in studying transcriptional regulation during brain development?

DLX1 antibody can reveal critical aspects of neurodevelopmental transcriptional regulation:

• DLX transcription factors are master regulators of the developing vertebrate brain, driving forebrain GABAergic neuronal differentiation

• ChIP-seq studies using DLX1 antibody can identify direct target genes and binding patterns throughout development

• Temporal analysis across developmental stages (e.g., E11.5, E13.5, E16.5) reveals dynamic binding patterns that change during neuronal differentiation

• Genome-wide binding analysis can identify chromatin states associated with DLX1 binding and regulatory element activity

• Comparison of wild-type and Dlx1/2-/- tissues can identify regions with differential histone post-translational modification signals, providing insights into the functional consequences of DLX1 binding

How does DLX1 interact with chromatin modifiers and what methods can detect these interactions?

Recent research has identified important interactions between DLX1 and chromatin modifiers:

• Tandem mass spectrometry following DLX1-FLAG immunoprecipitation has identified interactions with components of the Nucleosome Remodeling and Deacetylase (NuRD) complex

• Co-immunoprecipitation experiments with DLX1-FLAG and StrepII-tagged NuRD subunits in HEK-293T cells can validate these interactions

• DLX1 specifically interacts with RBBP4 and RBBP7 components of the NuRD complex

• These interactions are functionally important for decommissioning regulatory elements of genes such as Olig2, altering chromatin accessibility

• For optimal detection of these interactions, cellular fractionation to remove cytoplasmic proteins and treatment of nuclear lysates with Benzonase to minimize DNA/RNA-mediated interactions are recommended

What is the significance of DLX1 as a biomarker in cancer research?

DLX1 has emerged as a potential biomarker with significant implications for cancer research:

• DLX1 mRNA expression is significantly upregulated in lung adenocarcinoma (LUAD) compared to normal tissues

• High DLX1 expression or promoter methylation is associated with worse prognosis in LUAD patients

• DLX1 has been confirmed as an independent prognostic factor in LUAD through multivariate analysis

• In addition to LUAD, DLX1 expression is significantly higher in tumor tissues compared to normal tissues across multiple cancer types

• DLX1 may have diagnostic value, as demonstrated by ROC curve analysis and nomogram model analysis

How does DLX1 expression correlate with immune cell infiltration in tumor microenvironments?

Research has established important connections between DLX1 expression and immune cell infiltration:

• The level of multiple immune cell infiltration is significantly associated with DLX1 expression in LUAD

• Spearman correlation analysis can determine the relationship between DLX1 expression and immune cell infiltration

• This correlation may have implications for immunotherapy responses and patient outcomes

• The association between DLX1 and immune infiltration provides insights into potential mechanisms by which DLX1 influences tumor progression beyond its direct effects on cancer cells

What signaling pathways are associated with DLX1 in cancer progression?

DLX1 is involved in multiple signaling pathways relevant to cancer progression:

• In LUAD, genes in the high DLX1 expression group are mainly enriched in cell cycle checkpoint, DNA replication, DNA repair, Fceri-mediated MAPK activation, TP53 activity regulation, and MET activation of PTK2-regulated signaling pathways

• In ovarian cancer, DLX1 promotes aggressiveness by activating the TGF-β/SMAD4 signaling pathway

• In prostate cancer, DLX1 functions as a β-catenin binding protein, and aberrant activation of β-catenin/TCF signaling promotes growth, migration, and invasion

• These pathways can be analyzed through GO and KEGG pathway analysis of genes differentially expressed between high and low DLX1 expression groups

What methodological approaches can validate DLX1 as a therapeutic target in cancer?

To validate DLX1 as a therapeutic target in cancer, researchers should consider:

• Knockdown experiments: siRNA or shRNA targeting DLX1 has been shown to inhibit proliferation, migration, and invasion of LUAD cells

• Functional assays: Cell proliferation, migration, and invasion assays following DLX1 manipulation

• Prognostic analysis: Kaplan-Meier survival curves, univariate and multivariate Cox regression analyses to evaluate the association between DLX1 expression and patient outcomes

• Logistic regression analysis to evaluate the relationship between DLX1 expression levels and clinicopathologic characteristics

• Analysis of DLX1 mutation rates and their impact on prognosis using tools like cBioPortal

• Evaluation of DNA methylation levels of DLX1 and the prognostic value of CpG islands using methods such as MethSurv

How can researchers address inconsistent results when using DLX1 antibody across different applications?

When faced with inconsistent results:

• Verify antibody specificity using knockout controls when possible

• Consider that DLX1 may have overlapping functions with other DLX family members (DLX2, DLX5, DLX6), which may complicate interpretation of results

• Optimize antibody concentration through titration experiments specific to each application

• Ensure proper antigen retrieval methods for IHC applications, testing both recommended methods (TE buffer pH 9.0 and citrate buffer pH 6.0)

• Confirm appropriate positive controls are being used (human brain tissue, A375 cells, HeLa cells for WB; mouse brain tissue for IHC)