mpst-4 Antibody

Here’s a structured collection of FAQs tailored for academic researchers working with MPST-4 antibodies, integrating technical depth, methodological guidance, and evidence-based insights:

What are the validated applications for MPST-4 antibody in experimental workflows?

MPST-4 antibody (IgG1 κ mouse monoclonal) is optimized for:

• Western blot (WB): Detects MPST (~33 kDa) in mouse, rat, and human lysates under denaturing conditions .

• Immunohistochemistry (IHC): Requires antigen retrieval for formalin-fixed, paraffin-embedded tissues .

• Immunofluorescence (IF): Suitable for detecting cytoplasmic MPST in frozen sections .

• ELISA: Use non-conjugated forms for direct antigen quantification .

Methodological Note:

• For WB, reduce sample processing variability by using fresh lysates with protease inhibitors and validating with MPST-knockout controls .

• For IHC, optimize blocking conditions (e.g., 5% BSA in PBS) to minimize background .

How to confirm MPST-4 antibody specificity in complex biological samples?

Follow a three-step validation framework:

• Genetic controls: Compare wild-type vs. MPST-knockout cell lines or tissues .

• Orthogonal validation: Use siRNA-mediated MPST knockdown alongside antibody staining .

• Cross-reactivity profiling: Test against related sulfurtransferases (e.g., TST) to rule off-target binding .

Example data:

How to resolve discrepancies in MPST-4 antibody performance across assays?

Common issues and solutions:

Case Study:
In a study of MPST’s role in cysteine metabolism, inconsistent WB results were traced to variable β-mercaptoethanol concentrations during sample prep. Standardizing reducing conditions resolved the issue .

How to design a multiplex assay combining MPST-4 antibody with metabolic probes?

Workflow:

• Fixation: Use 4% PFA for 15 min to preserve MPST epitopes while retaining small-molecule probe integrity .

• Sequential staining:

  • Perform IF for MPST (Cy3-conjugated antibody).

  • Quench autofluorescence with 0.1% Sudan Black.

  • Incubate with tetrazine-conjugated metabolic probes (e.g., cysteine analogs) .

• Imaging: Use spectral unmixing to separate signals (e.g., Cy3 vs. Alexa Fluor 647 channels) .

Critical Consideration:
Validate probe-antibody compatibility via control experiments omitting either component .

What statistical approaches are robust for analyzing MPST-4 antibody-derived datasets?

Adopt finite mixture models to distinguish antibody-positive/negative populations in heterogeneous samples:

• Model: Skew-Normal distributions to account for asymmetric signal intensities .

• Validation: Compare AIC/BIC values across 2-4 component models .

Technical Notes

• Batch Consistency: Recombinant MPST-4 antibodies show ≤15% inter-batch variability in affinity measurements .

• Epitope Mapping: MPST-4 targets residues 89-103 in the N-terminal rhodanese domain (UniProt: P25325) .