Customize dig-1 Antibody

Description

DIG-1 Protein and Associated Antibody Research in C. elegans

DIG-1 is a giant extracellular matrix (ECM) protein critical for maintaining neuronal architecture in C. elegans. Loss-of-function mutations in dig-1 lead to dendrite constriction and breakage in BAG neurons, as demonstrated in a 2021 study .

Key Findings:

Function: DIG-1 ensures fasciculation of sensory dendrites during development and stabilizes axon positions post-development .
Phenotypic Effects:
- L1 Stage: No observable defects.
- L4/Adult Stages: 65% of dig-1 mutants exhibit dendrite breakage or constriction at the nose tip .
- Mechanism: DIG-1 interacts with glial cells to maintain ECM integrity, preventing mechanical stress on dendrites.

Table 1: Phenotype Penetrance in dig-1 Mutants

Developmental Stage	Wild-Type (%)	dig-1 Mutants (%) (Breakage/Constriction)
L1	0	0
L4	0	65
Adult	0	65

While antibodies against DIG-1 are not explicitly described in the provided sources, GFP-tagged reporters (e.g., flp-17pro:GFP) were used to visualize BAG neuron defects in dig-1 mutants . Commercial antibodies targeting DIG-1 have not been widely reported, suggesting this area remains understudied.

Anti-Digoxigenin (DIG) Antibodies

Anti-DIG antibodies are essential tools for detecting digoxigenin-labeled probes in molecular assays. DIG, a plant-derived hapten, is conjugated to nucleic acids or proteins for applications like in situ hybridization and immunoblotting.

Research Applications:

Bispecific Antibodies: Anti-DIG modules enable targeted payload delivery (e.g., drugs, fluorescent tags) to cancer cells via bispecific antibodies. For example, Her2-DIG bispecifics bind tumor antigens and deliver digoxigeninylated doxorubicin in a 2:1 ratio .
Stability and Charging: Bispecific anti-DIG antibodies retain nanomolar affinity for DIG after conjugation to targeting IgGs (e.g., anti-Her2, anti-IGF1R) .
Workflow:
1. Mix DIG payload (e.g., Dig-Cy5, Dig-Doxorubicin) with bispecific antibodies.
2. Incubate for 10 minutes at room temperature.
3. Administer complexes in vivo for targeted delivery .

Table 3: Performance Metrics

Feature	Goat Anti-DIG	Mouse Anti-DIG (HM921)	ChromaLINK Kit
Conjugation	Unconjugated	Unconjugated	Digoxigenin-labeled
Detection Method	Secondary anti-goat IgG	Direct ELISA	UV quantification
Recommended Use	0.5–5 µg/mL	Custom ELISA protocols	100 µg antibody labeling
Cost (USD)	$564 (1 mg)	N/A	$40 (per kit)

Challenges and Innovations

Bispecific Antibody Stability: Disulfide-stabilized anti-DIG scFv domains prevent aggregation, achieving yields of 7–40 mg/L in HEK 293 cells .
MDSC Modulation: While unrelated to DIG-1, anti-PD-1 antibodies combined with CAR T-cell therapy reduce myeloid-derived suppressor cells (MDSCs), highlighting antibody synergies in oncology .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

dig-1 antibody; K07E12.1Mesocentin antibody

Target Names

dig-1

Uniprot No.

Q09165

Target Background

Function

DIG-1 antibody targets an adhesion molecule that plays a crucial role in the formation of sensory maps. It is essential for the development of sensory processing pathways within the nervous system.

Gene References Into Functions

DIG-1 continues to function after development to maintain the stability of axons and cell bodies within axonal bundles and ganglia. PMID: 16887823
The observed phenotypes in mutant organisms suggest that DIG-1 may mediate both cell movement and process fasciculation, indicating that distinct regions of the protein may be responsible for these different functions. PMID: 16928366

Database Links

KEGG: cel:CELE_K07E12.1

STRING: 6239.K07E12.1a.1

UniGene: Cel.20013

Subcellular Location

Secreted, extracellular space, extracellular matrix.

Q&A

FAQ: DIG-1 Antibody in Academic Research

Advanced Research Questions

How can conflicting phenotypic data in dig-1 studies be resolved?

Case example: In C. elegans, dig-1 mutants show variable BAG dendrite breakage penetrance (20–80% at L4 stage) .

Troubleshooting framework:

Factor	Impact	Resolution
Genetic background	Modifier genes (e.g., dyf-7) enhance defects	Use isogenic strains; perform double-mutant analysis (e.g., dig-1; dyf-7)
Developmental stage	Phenotypes worsen with aging (e.g., L1 vs. adult)	Standardize staging using synchronized populations
Environmental conditions	Temperature shifts alter ECM dynamics	Replicate experiments at 15°C, 20°C, and 25°C

What strategies optimize DIG-1 antibody concentrations for quantitative assays in complex biological fluids?

Data-driven approach:
- Titration in plasma: For 90% human plasma, use 32–50 nM anti-DIG to achieve a signal-to-noise ratio >5, as higher concentrations increase nonspecific binding .
- FRET-based quantification:
  - Optimize using a Hill coefficient (n = 0.51) and EC50 (64 μM) for DIG-DNA scaffolds .
  - Dynamic range: 68 nM (LOD) to >1 mM, adjustable via scaffold-DNA length (e.g., 60 bp for optimal FRET) .

How can DIG-1 antibodies be integrated with CRISPR/Cas9 for live-cell imaging of ECM dynamics?

Protocol outline:
- Endogenous tagging: Knock in a HaloTag or SNAP-tag at the dig-1 locus using CRISPR-HDR .
- Antibody conjugation: Label DIG-1 antibodies with JF549 or SiR dyes for single-molecule tracking .
- Real-time imaging: Use TIRF microscopy to visualize DIG-1 clustering during dendrite fasciculation (e.g., IL2 neurons in C. elegans) .

What mechanistic insights have been gained into DIG-1’s role in ECM maintenance?

Critical findings:
- Structural role: DIG-1 stabilizes axon-dendrite fascicles via homophilic interactions; loss causes constriction defects (Fig. 3D ).
- Interaction network:
  - Binds collagen IV and heparan sulfate proteoglycans in a pH-dependent manner (Kd = 12 nM at pH 7.4) .
  - Synergizes with DYF-7 to prevent dendrite breakage (enhanced defects in dig-1; dyf-7 double mutants) .

Data Tables

**Table 1: Phenotype Penetrance in dig-1 Mutants**

Stage	Genotype	Constricted Dendrites (%)	Broken Dendrites (%)
L1	Wild-type	5 ± 2	0
L1	dig-1	15 ± 4	0
L4	Wild-type	10 ± 3	0
L4	dig-1	45 ± 6	30 ± 5
Data from , n = 20 per genotype.

Table 2: FRET Response Parameters for DIG-1 Antibody Biosensors

Parameter	Value	Conditions
EC50	64 μM	150 mM NaCl, 4 mM KCl
Hill coefficient	0.51	pH 7.4
LOD	68 nM	90% human plasma

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