Spink2 Antibody
Description
Introduction to Spink2 Antibody
The Spink2 Antibody is a research tool designed to detect the serine protease inhibitor Kazal type 2 (SPINK2) protein, which plays a critical role in regulating protease activity in cellular environments . SPINK2 is part of the SPINK family, known for its ability to inhibit serine proteases through high-affinity binding, acting as a temporary inhibitor . The antibody is primarily utilized in immunohistochemistry (IHC), western blotting, and flow cytometry to study SPINK2 expression in tissues and cells, particularly in hematopoietic malignancies .
Role in Hematopoietic Research
Key Findings:
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SPINK2 is highly expressed in hematopoietic stem and progenitor cells (HSPCs), where it modulates intercellular communication within the bone marrow niche .
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Its interaction with proteases like PRSS2 and PRSS57 suggests a regulatory mechanism in stem cell maintenance .
Data Table: SPINK2 Expression in Hematopoietic Cells
| Cell Type | SPINK2 Expression | Target Proteases | Reference |
|---|---|---|---|
| Hematopoietic Stem Cells | High | PRSS2, PRSS57 | |
| Mobilized CD34+ Cells | Elevated | PRSS2, PRSS57 |
Clinical Prognostic Value in Acute Myeloid Leukemia (AML)
Key Findings:
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High SPINK2 protein expression, detected via IHC, is an independent adverse prognostic biomarker in AML .
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Patients with elevated SPINK2 levels exhibit lower complete remission (CR) rates (34% vs. 52%) and higher non-remission (NR1) rates (45% vs. 28%) .
Data Table: Prognostic Outcomes in AML Patients
Therapeutic Potential
Key Developments:
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SPINK2 inhibition enhances immune response by upregulating ALCAM, a T-cell activation promoter .
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A small-molecule inhibitor (SMI) targeting SPINK2 shows promise in reducing ferroptosis resistance and improving treatment efficacy in AML .
Data Table: SPINK2 Inhibition Effects
| Parameter | Baseline | Post-Inhibition | Reference |
|---|---|---|---|
| SLC7A11 Expression | 1.0 | 0.6 | |
| Erastin Sensitivity | 50% | 80% | |
| ALCAM Expression | 1.0 | 2.5 |
Challenges and Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Spink2 is a potent inhibitor of acrosin, an enzyme essential for normal spermiogenesis. Its role is to prevent premature activation of proacrosin and other proteases, thus safeguarding against the cascade of events leading to spermiogenesis defects. It may also play a regulatory role in serine protease-dependent germ cell apoptosis. Additionally, Spink2 exhibits inhibitory activity against trypsin.
- Overexpression of different variants of Spink2 in D407 tissue culture cells enhances their sensitivity to the apoptosis-inducing agent staurosporine, aligning with the observed differential susceptibility between the DBA/2J and BALB/cByJ strains. PMID: 24699552
- SPINK2 is crucial for maintaining normal spermatogenesis and potentially regulates serine protease-mediated apoptosis in male germ cells. PMID: 21705336
Q&A
Basic Research Questions
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What is SPINK2 and what cellular functions does it regulate?
SPINK2 (Serine Protease Inhibitor Kazal type 2) functions as a serine protease inhibitor that regulates proteolytic processes. It acts as a strong inhibitor of acrosin and is required for normal spermiogenesis, preventing premature activation of proacrosin and other proteases. SPINK2 inhibits the cascade of events that can lead to spermiogenesis defects . Additionally, SPINK2 may be involved in the regulation of serine protease-dependent germ cell apoptosis and can inhibit trypsin .
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Which applications are most suitable for SPINK2 antibody detection?
SPINK2 antibodies have been validated for multiple research applications, including Western blot (WB), enzyme-linked immunosorbent assay (ELISA), immunofluorescence (IF), and immunohistochemistry on paraffin-embedded tissues (IHC-P) . IHC-P appears particularly reliable for detecting SPINK2 in tissue samples and has been extensively used in prognostic studies of acute myeloid leukemia (AML) . Western blotting can detect SPINK2 protein bands of less than 17 kDa weight, likely corresponding to oligomeric complexes .
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How should SPINK2 antibody specificity be validated?
Validation of SPINK2 antibody specificity should involve multiple approaches:
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Western blot analysis using cells overexpressing SPINK2 alongside non-transfected controls
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Immunofluorescence experiments comparing transfected versus non-transfected cells
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Using tagged versions of SPINK2 (such as DDK-tagged) that can be detected with anti-tag antibodies to confirm specificity
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Including appropriate negative controls (omitting primary antibody)
Research has demonstrated that specific SPINK2 antibodies recognize three bands of less than 17 kDa in Western blots from SPINK2-overexpressing cells, with no bands appearing in non-transfected cells .
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What tissue types show the highest expression of SPINK2 for antibody detection?
SPINK2 exhibits tissue-specific expression patterns, with the highest expression found in the testis . For antibody detection studies, human and mouse testis tissues have been successfully used to evaluate SPINK2 localization using immunohistochemistry and immunofluorescence . Additionally, SPINK2 expression has been detected in bone marrow specimens from AML patients, making this tissue type relevant for clinical studies .
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What dilution ranges are typically effective for SPINK2 antibody applications?
Based on published methodologies, effective dilution ranges for SPINK2 antibodies vary by application:
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For IHC-P: 1/500 dilution has been reported as effective for staining in human testis tissue
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For Western blot: Optimal dilutions typically range from 1:1000 to 1:5000, depending on the specific antibody
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For immunofluorescence: Dilutions around 1:200 have been reported in cell-based assays
It's recommended to perform titration experiments to determine optimal concentrations for each specific antibody and experimental system .
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