Customize dhp-2 Antibody

Description

Definition and Target Specificity

Antibodies targeting dihydropyridine (DHP)-associated proteins are specialized tools used to study voltage-gated calcium channels (VGCCs), particularly L-type calcium channels (LTCCs). These antibodies recognize epitopes on α-subunits of LTCCs (e.g., CaV1.2, encoded by CACNA1C), which are critical for calcium signaling in cardiac, skeletal, and neuronal tissues . DHPs, such as nifedipine and Bay k 8644, bind to these channels to modulate their activity .

Antibody Architecture

Heavy and Light Chains: Composed of two heavy chains (~50 kDa) and two light chains (~25 kDa) forming a Y-shaped structure with Fab (antigen-binding) and Fc (effector) regions .
Epitope Recognition:
- ACC-003: Targets residues 848–865 of CaV1.2 (intracellular loop between domains II and III) .
- IIID5E1: Binds the α-subunit of skeletal muscle DHPR (CaV1.1) .

Target Protein Features

Feature	Ca<sub>V</sub>1.2 (ACC-003)	DHPR α-Subunit (IIID5E1)
Molecular Weight	~220 kDa	~170 kDa
Localization	Cardiac myocytes, Purkinje cells	Skeletal muscle T-tubules
Key Domains	Pore-forming α<sub>1</sub>-subunit	Voltage-sensing domain IVS6

Common Use Cases

Western Blot: Detects CaV1.2 in rat brain membranes (1:200 dilution) .
Immunohistochemistry: Labels Purkinje cells in mouse cerebellum (ACC-003) and human skeletal muscle (IIID5E1) .
Flow Cytometry: HPa2 (DHIC2-2B4) targets pancreatic α-cells (PE conjugate, 2–5 µg/mL) .

Validation Data

Antibody	Species Reactivity	Key Findings	Source
ACC-003	Human, Rat, Mouse	Co-localizes with β-arrestin in cardiomyocytes	Alomone Labs
IIID5E1	Human, Mouse, Rabbit	Confirmed in HEK-293 and mouse heart lysates	DSHB
HPa2	Human	Specific to pancreatic α-cells (cell surface)	Bio-Techne

Mechanistic Insights

DHP Binding: Agonists (e.g., Bay k 8644) and antagonists (e.g., nimodipine) interact with LTCCs via H-bonds to TyrIIIS6 and GlnIIIS5, stabilizing open or closed states .
Antibody-Drug Interactions: ACC-003 blocks CaV1.2 function, validated in Xenopus oocytes .

Clinical Correlations

Cardiac Disease: CaV1.2 dysregulation linked to arrhythmias .
Neuromuscular Disorders: DHPR mutations cause malignant hyperthermia .

Comparative Analysis of Key Antibodies

Parameter	ACC-003	IIID5E1	HPa2
Host Species	Guinea Pig	Mouse	Mouse
Isotype	Polyclonal	IgG1	IgG1
Applications	WB, IHC, IP	WB, IHC, IP	Flow Cytometry, IHC
Commercial Source	Alomone Labs	DSHB	Bio-Techne

Future Directions

Therapeutic Development: Antibodies like ACC-003 could aid in designing LTCC-targeted drugs for hypertension .
Diagnostic Tools: HPa2 may improve α-cell dysfunction detection in diabetes .

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

dhp-2 antibody; C47E12.8Dihydropyrimidinase 2 antibody; EC 3.5.2.2 antibody; CeCRMP/DHP-2 antibody; UlipA antibody

Target Names

dhp-2

Uniprot No.

Q18677

Q&A

Here’s a structured FAQ collection for researchers working with the DHP-2 antibody, tailored to academic research scenarios and informed by experimental methodologies from peer-reviewed practices and patents:

Advanced Research Questions

How do I resolve contradictions in DHP-2 localization data between studies?

Troubleshooting matrix:

Issue	Potential Cause	Resolution
Nuclear vs. cytoplasmic	Cross-reactivity with splice variants	Perform siRNA rescue experiments
Discrepant IHC vs. WB	Epitope masking in fixed tissues	Use antigen retrieval (e.g., citrate buffer, pH 6.0)

What strategies improve DHP-2 detection in low-abundance protein complexes?

Enrichment protocols:
- Immunoprecipitation (IP): Pre-clear lysates with protein A/G beads, then incubate with DHP-2 antibody (4°C overnight) .
- Proximity ligation assays (PLA): Amplify signals for protein-protein interactions (e.g., Duolink® kits) .
- Mass spectrometry: Combine IP with LC-MS/MS for interactome profiling (see Figure 14A-14D in ).

How do I design a multiplex assay integrating DHP-2 with other biomarkers?

Workflow:
- Panel design: Prioritize high-expression biomarkers for dimmer fluorophores (e.g., FITC) .
- Competition testing: Pre-mix DHP-2 with other primary antibodies to assess steric hindrance.
- Validation: Use spike-in samples with known biomarker combinations .

Methodological Insights from Recent Studies

COX-2 interaction analysis (from ):
- Photoaffinity labeling + LC-MS/MS identified 1034 enriched proteins in Jurkat/K562 cells, with statistical filtering (FDR < 0.05).
- Critical parameters: 6M urea washes reduced nonspecific binding in streptavidin pulldowns (Figure 19 in ).
Dose-response profiling:

Compound IC₅₀ (Jurkat cells) COX-2 Inhibition
Photo-naproxen 18.3 µM 92% at 50 µM
Photo-celecoxib 9.7 µM 88% at 25 µM
Data derived from ELISA and MTT assays (Figures 16A-16D, 17D in ).

Compound	IC₅₀ (Jurkat cells)	COX-2 Inhibition
Photo-naproxen	18.3 µM	92% at 50 µM
Photo-celecoxib	9.7 µM	88% at 25 µM
Data derived from ELISA and MTT assays (Figures 16A-16D, 17D in ).

Data Contradiction Analysis Framework

Case example: If DHP-2 shows inconsistent binding in replicate experiments:
- Technical variability: Check antibody storage (avoid freeze-thaw cycles >3x) .
- Biological variability: Use isogenic cell lines or primary cells from ≥3 donors.
- Statistical rigor: Apply Benjamini-Hochberg correction for high-throughput datasets .

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dhp-2 Antibody