Customize MSRB2 Antibody

Description

Biological Functions of MSRB2

MSRB2 is a mitochondrial enzyme with dual roles in oxidative stress response and cellular homeostasis:

Key Functions

Antioxidant Activity: Reduces methionine-R-sulfoxides, counteracting ROS-induced protein damage .
Mitophagy Regulation: Activates Parkin by reducing its methionine oxidation, enabling ubiquitination of damaged mitochondria and LC3-mediated autophagosomal clearance .
Cytokinetic Control: Modulates actin dynamics during cell division by countering MICAL1-mediated oxidation, ensuring midbody stability .

Pathophysiological Roles

Diabetes: Upregulated in diabetic platelets, enhancing mitophagy to suppress apoptosis .
Neurodegeneration: Reduced MSRB2 levels in Parkinson’s disease correlate with impaired mitophagy .
Cardioprotection: Mitigates diabetic cardiomyopathy by preserving mitochondrial energy metabolism .

3.1. Mitophagy and Apoptosis (2019)

Mechanism: MSRB2 released from damaged mitochondria reduces Parkin MetO, enabling Parkin-LC3 interaction for mitophagy initiation .
Knockout Effects: Platelet-specific MSRB2 deletion increases ROS, mitochondrial membrane depolarization, and apoptosis .
Clinical Relevance: Elevated MSRB2 in diabetes enhances platelet survival; reduced levels in Parkinson’s exacerbate mitochondrial dysfunction .

3.2. Diabetic Cardiomyopathy (2024)

Protective Role: MSRB2 upregulation in diabetic hearts preserves mitochondrial function via LC3 activation and Parkin interaction, reducing cardiomyopathy progression .

3.3. Cytokinesis Regulation (2020)

Actin Dynamics: MSRB2 depletion accelerates abscission by destabilizing intercellular bridge actin, while overexpression delays it via Aurora B-dependent pathways .
ESCRT-III Recruitment: MSRB2 counteracts MICAL1 to regulate F-actin levels, ensuring proper ESCRT-III localization for abscission .

Comparative Antibody Performance

Feature	Abcam (ab229940)	Proteintech (17629-1-AP)	Novus (NBP1-86594)
Applications	WB, IHC-P	WB, IHC, ELISA	IHC, WB
Tissue Reactivity	Human, Mouse	Human, Mouse, Rat	Human
Dilution Range	WB: 1/1000; IHC: 1/100	WB: 1:500–1:1000	WB: 0.04–0.4 µg/mL
Validation	In-house + user data	Published studies	Orthogonal validation

Clinical and Therapeutic Implications

Diabetes Management: Targeting MSRB2 could modulate platelet apoptosis and cardiovascular outcomes .
Neuroprotection: Enhancing MSRB2 activity may mitigate mitochondrial dysfunction in neurodegenerative diseases .
Cancer: Aberrant MSRB2 expression influences cell division fidelity, with implications for genomic stability .

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

MSRB2 antibody; At4g21860 antibody; T8O5.70 antibody; Peptide methionine sulfoxide reductase B2 antibody; chloroplastic antibody; AtMSRB2 antibody; EC 1.8.4.12 antibody; Peptide-methionine antibody; R)-S-oxide reductase antibody

Target Names

MSRB2

Uniprot No.

Q9C5C8

Target Background

Function

This antibody targets MSRB2, an enzyme that catalyzes the reduction of methionine sulfoxide (MetSO) to methionine in proteins. Specifically, it reduces the MetSO R-enantiomer. MSRB2 plays a protective role against oxidative stress by restoring activity to proteins that have been inactivated by methionine oxidation. It may also play a crucial function in association with MSRB1 in maintaining vegetative growth during environmental constraints, through the preservation of photosynthetic antennae. Notably, MSRB1 and MSRB2 account for the majority of the leaf peptide MSR capacity.

Gene References Into Functions

Research suggests that plastidial MSRBs have a crucial role in maintaining the vegetative growth of plants during environmental constraints, achieved through their involvement in the preservation of photosynthetic antennae. PMID: 19874542

Database Links

KEGG: ath:AT4G21860

STRING: 3702.AT4G21860.1

UniGene: At.24358

Protein Families

MsrB Met sulfoxide reductase family

Subcellular Location

Plastid, chloroplast.

Tissue Specificity

Expressed in stems, young leaves, floral buds and flowers. Expressed at low levels in roots, mature leaves and siliques (at protein level).

Q&A

Basic Research Questions

How to validate MSRB2 antibody specificity in Western blot?

Method: Use tissue/cell lysates with confirmed MSRB2 expression (e.g., human kidney, mouse brain) alongside knockout controls.
Key controls:
- Pre-adsorption with immunogen peptide ( ).
- Compare observed molecular weight (~19 kDa) to calculated weight (20 kDa) to detect truncation or degradation ( ).
- Validate in species-specific models (human, mouse, rat) using tissues listed in validation galleries ( ).

What are optimal protocols for MSRB2 detection in mitochondrial vs. cytoplasmic compartments?

Mitochondrial enrichment: Pre-fractionate samples using differential centrifugation.
IHC optimization: Use TE buffer (pH 9.0) or citrate buffer (pH 6.0) for antigen retrieval to improve mitochondrial epitope exposure ( ).
Cytoplasmic detection: For non-mitochondrial pools (e.g., cytokinetic studies), combine antibody staining with GFP-tagged MSRB2 constructs to resolve localization ambiguities ( ).

How to troubleshoot non-specific bands in Western blot?

Likely causes: Cross-reactivity with other methionine sulfoxide reductases (e.g., MSRB1/MSRB3).
Solutions:
- Titrate antibody concentration (start at 1:500 dilution) ( ).
- Include knockout lysates to identify off-target bands ( ).
- Use reducing conditions to prevent disulfide-linked aggregates ( ).

Advanced Research Questions

How to design experiments analyzing MSRB2’s role in redox-dependent cytoskeletal regulation?

Functional assays:
- Pair MSRB2 antibodies with in vitro actin oxidation assays using MICAL1-treated actin and measure reduction efficiency via subtilisin A cleavage ( ).
- Monitor cytokinetic abscission timing in MSRB2-depleted cells using live imaging (e.g., CHMP4B-EGFP reporters) ( ).
Key controls: Co-deplete MICAL1 to reverse accelerated abscission phenotypes ( ).

What methods resolve contradictions in MSRB2 localization studies?

Conflict: Mitochondrial vs. cytoplasmic pools reported in PNAS ( ) vs. vendor datasheets ( ).
Resolution strategies:
- Use subcellular fractionation + Western blotting (mitochondrial matrix markers: COX IV; cytoplasmic markers: GAPDH).
- Combine antibody staining with MSRB2-GFP constructs to distinguish overexpression artifacts ( ).

How to assess MSRB2’s enzymatic activity in cellular contexts?

Approach:
- Measure methionine-R-sulfoxide reduction rates using LC-MS in MSRB2-KO cells rescued with wild-type vs. catalytic cysteine mutants.
- Correlate activity with mitochondrial ROS levels via MitoSOX™ Red staining ( ).

Methodological Reference Table

Application	Critical Parameters	Supporting Data Sources
Western Blot	Lysate preparation with protease/phosphatase inhibitors; 10–15% gradient gels	(tested tissues), (cross-reactivity)
IHC	Antigen retrieval buffer pH optimization; validation in FFPE vs. frozen sections	(human kidney), (mouse models)
Functional Studies	Co-depletion with Aurora B/ANCHR; actin polymerization assays	(cytokinetic abscission), (MICAL1/MsrB2 interaction)

Data Discrepancy Analysis Framework

For conflicting results (e.g., antibody reactivity across studies):

Compare immunogens: Full-length protein ( ) vs. peptide fragments ( ) may yield varying epitope access.
Assay context: Mitochondrial enrichment protocols differ between studies ( vs. ).
Species specificity: Rabbit polyclonals ( ) vs. mouse monoclonals ( ) show distinct cross-reactivity profiles.

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MSRB2 Antibody