Customize pink-1 Antibody

Description

Introduction to PINK1 Antibody

PINK1 (PTEN-induced putative kinase 1) is a serine/threonine kinase critical for mitochondrial quality control and implicated in neurodegenerative diseases like Parkinson’s disease (PD) and cancer biology. PINK1 antibodies are essential research tools designed to detect, quantify, and study the subcellular localization, expression, and functional interactions of PINK1 in experimental models. These antibodies target specific epitopes across the PINK1 protein, enabling applications such as Western blotting (WB), immunocytochemistry (ICC), and immunohistochemistry (IHC).

Epitope Specificity and Validation

PINK1 antibodies are developed against distinct regions of the protein, including:

N-terminal mitochondrial targeting sequence (residues 1–50; NB100-493 )
Kinase domain (residues 175–250; BC100-494 )
Central region (residues 258–274; PA1-4515 )

Antibody Clone	Target Region	Applications	Key Findings
BC100-494	Residues 175–250	WB, ICC, IHC, IP	Detects endogenous PINK1 (~63 kDa) and cleaved forms (55 kDa, 42 kDa) .
NB100-493	N-terminal (1–50)	WB (~63 kDa)	Identifies unprocessed PINK1; detects murine and human isoforms .
PA1-4515	Residues 258–274	WB (HEK293T lysates)	Confirms PINK1 overexpression in transfected cells .
SMC-450	Unspecified epitope	ICC, WB (neuroblastoma cells, rat brain)	Localizes PINK1 to cytoplasm and mitochondria in human cell lines .

Species Reactivity

Validated in human, mouse, and rat models, with cross-reactivity confirmed in mitochondrial stress assays (e.g., CCCP/valinomycin treatment) .

Mitochondrial Dynamics and Parkinson’s Disease

Mechanism: PINK1 accumulates on depolarized mitochondria, forming dimers that recruit Parkin to initiate mitophagy .
- Phosphorylation of Parkin at Ser65 by PINK1 enhances its E3 ligase activity .
- Loss of PINK1 increases neuronal vulnerability to toxins (e.g., MPTP) .
Therapeutic Insight: Cytoplasmic PINK1 retains neuroprotective effects independent of mitochondrial localization .

Cancer Biology

Breast Cancer: High PINK1 expression correlates with histological grade (34.2% in grade I–II vs. 57.6% in grade III; p = 0.015) and promotes glycolysis in MDA-MB-231 cells .
Proteomic Impact: PINK1 deletion alters energy metabolism pathways, reducing glycolytic capacity (Seahorse XF analysis) .

Tauopathy and Cognitive Function

Tau Degradation: PINK1 overexpression reduces pathological tau accumulation in hippocampal neurons via autophagy-lysosomal pathways (ALP), rescuing synaptic deficits in tauopathy models .

Subcellular Localization

Dual Localization: PINK1 resides in both mitochondria and cytosol. Antibodies targeting the N-terminal region (e.g., NB100-493) detect precursor forms, while those against the kinase domain (e.g., BC100-494) identify processed isoforms .
Mitochondrial Stress: CCCP/valinomycin treatment enhances PINK1 antibody signal by stabilizing full-length PINK1 on depolarized mitochondria .

Validation Challenges

Band Specificity: Multiple bands (48, 55, 63 kDa) may appear due to proteolytic processing; siRNA knockdown or knockout controls are critical .
Phospho-Specific Reagents: Novel phospho-ubiquitin (p-S65-Ub) antibodies monitor PINK1-Parkin signaling in mitophagy assays .

Diagnostic Potential

Phospho-specific antibodies (e.g., p-Thr257-PINK1) may serve as biomarkers for PD progression .
PINK1 expression in breast cancer tissues is proposed as a malignancy indicator .

Antibody Development Initiatives

The Michael J. Fox Foundation supports monoclonal antibody development to target phosphorylated Parkin (Ser65) and PINK1 (Thr257) for therapeutic screening .

Table 1: PINK1 Antibody Performance in Select Studies

Study Focus	Antibody Used	Key Outcome
Mitophagy initiation	BC100-494	Detected PINK1/Parkin colocalization in RPE cells (WT vs. dKO mice)
Tau degradation	Unspecified	PINK1 reduced soluble/insoluble tau by 40–60% in hTau mice (p < 0.0001)
Glycolysis regulation	Commercial PINK1 siRNA	Reduced glycolytic capacity in MDA-MB-231 cells (Seahorse XF)

Table 2: Clinical Implications of PINK1 Dysregulation

Disease Context	PINK1 Role	Therapeutic Target
Parkinson’s disease	Loss of PINK1 increases neuronal toxicity; cytoplasmic PINK1 retains function	Kinase activators, mitophagy enhancers
Breast cancer	Oncogenic properties via glycolysis promotion	PINK1 inhibitors

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

pink-1 antibody; EEED8.9 antibody; Serine/threonine-protein kinase pink-1 antibody; mitochondrial antibody; EC 2.7.11.1 antibody; PTEN-induced kinase 1 homolog antibody

Target Names

pink-1

Uniprot No.

Q09298

Target Background

Function

This antibody targets a protein that protects against mitochondrial dysfunction during cellular stress, potentially through the phosphorylation of mitochondrial proteins. It also plays a role in mitophagy.

Gene References Into Functions

The role of this protein is further illustrated by studies on manganese uptake and manganese-induced oxidative stress in the context of mutated *PDR1*, *PINK1*, and *DJR1.1* genes. See PMID: 24452053 for details.

Database Links

KEGG: cel:CELE_EEED8.9

STRING: 6239.EEED8.9.2

UniGene: Cel.15543

Protein Families

Protein kinase superfamily, Ser/Thr protein kinase family

Subcellular Location

Mitochondrion.

Q&A

Here’s a structured collection of FAQs tailored for academic researchers working with PINK1 antibodies, based on technical insights from peer-reviewed studies and methodological data:

Advanced Research Questions

How does mitochondrial membrane potential affect PINK1 antibody detection?

Key Findings:
- Depolarized mitochondria (via CCCP/valinomycin) stabilize full-length PINK1 on the outer membrane, enhancing detection .
- Under normal conditions, PINK1 is cleaved to Δ2 (~52 kDa) and rapidly degraded; use proteasome inhibitors (epoxomicin) to enrich Δ2 .

What methods resolve contradictions in PINK1 localization studies?

Contradiction: Some studies report cytosolic PINK1, while others show mitochondrial association .
Resolution:
- Subcellular fractionation: Validate mitochondrial isolation purity (cytochrome c oxidase IV as marker) .
- Protease protection assay: Treat mitochondria with proteinase K to confirm membrane integration .
- Time-course experiments: PINK1 accumulates on depolarized mitochondria over 3–6 hrs .

How do post-translational modifications impact PINK1 antibody performance?

Critical Factors:
- Ubiquitination: Anti-PINK1 antibodies may detect polyubiquitinated smears (~70–150 kDa) under proteasome inhibition (MG132) .
- Phosphorylation: Use Phos-tag gels to separate phosphorylated species (e.g., Ser228/Ser402) .

Data Interpretation Challenges

Why do some antibodies fail to detect PINK1 in Lewy bodies?

Issue: Epitope masking by aggregated α-synuclein .
Solutions:
- Antigen retrieval: Autoclave sections in citrate buffer (pH 6.0) .
- Combine with proteinase K treatment (10 μg/mL, 5 min) .

How to address nonspecific bands in PINK1 Western blots?

Troubleshooting:

Band Size (kDa)	Likely Cause	Mitigation
~75–85	HSP90 cross-reactivity	Pre-clear lysates with HSP90 inhibitors (e.g., geldanamycin)
~45–50	PINK1 cleavage products	Use fresh lysates + protease inhibitors (e.g., PMSF, leupeptin)

Experimental Design Considerations

What controls are essential for PINK1 loss-of-function studies?

Required Controls:
- Rescue experiments: Re-express WT or kinase-dead (KD) PINK1 in knockout models .
- Mitochondrial stress controls: Include CCCP-treated cells to confirm Parkin recruitment .
- Antibody validation: Compare at least two independent antibodies (e.g., Novus BC100-494 vs. SMC-450) .

How to model PINK1-associated Parkinson’s disease in vitro?

Model System:
- Use patient-derived fibroblasts with heterozygous PINK1 mutations (e.g., G309D) .
- Differentiate SH-SY5Y cells into dopaminergic neurons (retinoic acid + BDNF), then induce mitochondrial stress (rotenone/MPP+) .

Quick Inquiry

Personal Email Detected

Please use an institutional or corporate email address for inquiries. Personal email accounts ( such as Gmail, Yahoo, and Outlook) are not accepted. *

Name*

Email*

Phone

Country

Source

Quantity&Size*

Product Name

Message

pink-1 Antibody