Customize CLH1 Antibody

Description

Definition and Biological Context

The CLH1 antibody is designed to detect and bind to CrCLH1, a 322-amino-acid enzyme expressed in Escherichia coli and immobilized for industrial biocatalysis. CrCLH1 catalyzes the hydrolysis of chlorophyll-a into chlorophyllide and phytol, with applications in producing chlorophyll derivatives . The antibody specifically recognizes epitopes such as the histidine (His) tag fused to the recombinant enzyme, enabling its purification and immobilization .

Enzyme Characteristics

Molecular Weight: ~35 kDa (predicted for recombinant CrCLH1 with tags) .
Tags: N-terminal T7 tag and C-terminal His tag for expression and detection .
Activity: Hydrolyzes chlorophyll-a optimally at pH 5.0 and 50–60°C .

Antibody Features

Target: His tag on recombinant CrCLH1.
Immobilization Method: Covalent binding to (3-aminopropyl) triethoxysilane (APTES)-coated magnetic iron oxide nanoparticles (MIONPs) .

Immobilization Efficiency and Activity

Immobilized CrCLH1 demonstrated enhanced stability and reusability compared to the free enzyme:

Parameter	Free CrCLH1	Immobilized CrCLH1
Immobilization Yield	N/A	98.99 ± 0.91%
Enzymatic Activity (U/g)	186 ± 22 (specific)	722.3 ± 50.3
Residual Activity (14 cycles)	N/A	64%

pH Stability: Retained activity in acidic environments (pH 3–5) .
Thermal Stability: Higher activity at 50–60°C than free enzyme .

Industrial Biocatalysis

Reusability: Retained 64% activity after 14 batch cycles, reducing production costs .
Scalability: Magnetic separation simplifies enzyme recovery for large-scale processes .

Chlorophyll Derivative Production

Converts chlorophyll-a to chlorophyllide-a, a precursor for natural food colorants and pharmaceuticals .

Future Directions

Optimization: Fine-tuning immobilization protocols to improve specific activity.
Diverse Substrates: Expanding catalytic activity to other chlorophyll types (e.g., chlorophyll-b).
Therapeutic Potential: Exploring CrCLH1’s role in photodynamic therapy or antioxidant production .

Product Specs

Buffer

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

Form

Liquid

Lead Time

14-16 weeks (Made-to-order)

Synonyms

CLH1 antibody; ATHCOR1 antibody; COR1 antibody; At1g19670 antibody; F6F9.28Chlorophyllase-1 antibody; AtCLH1 antibody; EC 3.1.1.14 antibody; Chlorophyll-chlorophyllido hydrolase 1 antibody; Chlase 1 antibody; Coronatine-induced protein 1 antibody; CORI1 antibody

Target Names

CLH1

Uniprot No.

O22527

Target Background

Function

This antibody targets Chlorophyllase 1 (CLH1), an enzyme that catalyzes the hydrolysis of ester bonds in chlorophyll, yielding chlorophyllide and phytol. It exhibits a preference for chlorophyll a. While not seemingly essential for chlorophyll degradation during senescence, CLH1 may regulate the balance between various plant defense pathways.

Gene References Into Functions

Research indicates a unique binary defense mechanism against insect herbivores involving chlorophyll and CLH1. [PMID: 25583926]
Findings suggest that AtCLH1 may be crucial in the early stages of leaf senescence, potentially facilitating the conversion of chlorophyll b to chlorophyll a. [PMID: 18349515]

Database Links

KEGG: ath:AT1G19670

STRING: 3702.AT1G19670.1

UniGene: At.19779

Protein Families

AB hydrolase superfamily, Lipase family

Subcellular Location

Cytoplasm, cytosol.

Tissue Specificity

Expressed in seedlings, leaves, flowers and siliques, but not in roots.

Q&A

The following FAQs address key considerations for researchers working with CLH1 Antibody in academic settings, structured to reflect scientific rigor and methodological depth. Questions are categorized into basic and advanced research tiers, with evidence-based answers incorporating proteomic analysis techniques and validation approaches.

Advanced Research Challenges

How to resolve contradictory CLH1 localization data between immunofluorescence and biochemical fractionation?

Troubleshooting Protocol:
- Perform simultaneous dual labeling with organelle-specific markers
- Validate fractionation purity using compartment-specific enzymes:
  - Mitochondrial: COX IV
  - Nuclear: Lamin B1
  - Cytoplasmic: GAPDH
- Assess antibody cross-reactivity via peptide blocking (≥5× molar excess)

What computational methods enhance CLH1 interaction partner identification in co-IP/MS studies?

Which statistical models best address batch effects in longitudinal CLH1 expression studies?

Advanced Normalization:
- Implement ComBat harmonization for multi-platform data
- Mixed-effects model framework:
  $Y_{ij} = μ + α_i + βX_{ij} + γZ_j + ε_{ij}$
  Where:
  - α = individual random effect
  - β = treatment fixed effect
  - γ = batch covariate

Data Conflict Resolution Matrix

Conflict Type	Validation Strategy	Required Replicates
Antibody lot variability	Parallel testing with NIST reference material	3 technical + 2 biological
Species cross-reactivity	Phylogenic informatics + recombinant ortholog testing	5 evolutionary nodes
Epitope masking	Limited proteolysis + epitope mapping via HDX-MS	2 independent methods

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CLH1 Antibody