PSMC1 Antibody

What is PSMC1 and what is its biological significance?

PSMC1, also known as 26S proteasome regulatory subunit 4 (PRS4) or P26s4, is a ubiquitously-expressed ATPase belonging to the triple-A family. It plays an essential role in the 19S regulatory particle of the 26S proteasome complex, which is critical for the controlled degradation of proteins in all cells . The 26S proteasome's proper function is vital for maintaining cellular homeostasis, and dysfunction of this system has been implicated in various neurodegenerative diseases due to the accumulation of abnormal proteins .

PSMC1 exists in a complex with PSMD2 and PSMD5, and research suggests it may serve as a protective reservoir against harmful effects resulting from suppression of the essential gene encoding PSMC2 . This protective function highlights PSMC1's importance in maintaining cellular integrity under stress conditions.

What types of PSMC1 antibodies are available for research applications?

Multiple types of PSMC1 antibodies are available for research purposes, varying in host species, clonality, epitope specificity, and applications:

These antibodies provide researchers with options for different experimental needs and model systems .

What are the standard applications for PSMC1 antibodies in laboratory research?

PSMC1 antibodies have been validated for multiple laboratory applications, including:

• Western Blotting (WB): For protein detection and quantification of PSMC1 expression levels in tissue/cell lysates .

• Immunohistochemistry (IHC): For visualization of PSMC1 distribution in paraffin-embedded tissue sections .

• Immunofluorescence (IF): For subcellular localization studies of PSMC1 .

• ELISA: For quantitative measurement of PSMC1 in solution .

• Immunocytochemistry (ICC): For examining PSMC1 expression in cultured cells .

The selection of the appropriate antibody and application depends on the specific research question, sample type, and desired level of quantitative or qualitative analysis.

How should I select an appropriate PSMC1 antibody for specific experimental applications?

When selecting a PSMC1 antibody for specific applications, researchers should consider:

• Epitope specificity: Different antibodies target various regions of the PSMC1 protein. For example, antibody ABIN1881692 targets amino acids 408-436 at the C-terminus , while ABIN7269584 recognizes a broader region (amino acids 77-440) . Select antibodies that target regions relevant to your research question, particularly if investigating specific domains or functional interactions.

• Host species compatibility: Consider experimental design, particularly for co-staining or multiplex applications. The available antibodies include mouse monoclonals (e.g., clone EF02/2E2) and rabbit polyclonals , allowing flexibility in experimental design.

• Cross-reactivity: Many PSMC1 antibodies show cross-reactivity across human, mouse, and rat samples , while some offer broader reactivity across multiple species including zebrafish, chicken, cow, guinea pig, horse, Xenopus laevis, pig, monkey, bat, and even Drosophila melanogaster . This wide cross-reactivity enables comparative studies across model organisms.

• Validated applications: Ensure the antibody is validated for your intended application. For instance, if performing immunohistochemistry on paraffin-embedded tissues, confirm the antibody has been validated for IHC(p) .

• Clonality considerations: Monoclonal antibodies like EF02/2E2 offer high specificity for a single epitope, reducing background and increasing reproducibility. Polyclonal antibodies may provide stronger signals by recognizing multiple epitopes but with potentially higher background.

What protocols should be optimized when using PSMC1 antibodies to investigate proteasome dysfunction in neurodegenerative disease models?

When using PSMC1 antibodies to study proteasome dysfunction in neurodegenerative disease models, consider these methodological optimizations:

• Sample preparation: Proteasome complexes are sensitive to degradation. Use fresh tissue samples or rapid fixation protocols to preserve the native state of the proteasomal complexes. For neuronal tissues, consider specialized fixation methods that maintain both tissue architecture and protein epitope accessibility.

• Co-immunoprecipitation studies: Since PSMC1 exists in a complex with PSMD2 and PSMD5 , co-immunoprecipitation protocols can help elucidate changes in these interactions under pathological conditions. Optimize lysis buffers that maintain protein-protein interactions while effectively disrupting cellular membranes.

• Functional assays: Beyond mere detection, consider using PSMC1 antibodies in combination with proteasome activity assays to correlate PSMC1 expression/localization with functional changes in proteasomal activity.

• Multiple markers approach: Given that incorrect function of the 26S proteasome has been implicated in neurodegenerative diseases , combine PSMC1 antibody staining with markers of protein aggregation (e.g., ubiquitin, p62) to correlate proteasome dysfunction with disease pathology.

• Controls: Include both positive controls (tissues known to express PSMC1) and negative controls (tissues from PSMC1 knockout models if available, or primary antibody omission controls) to validate specificity in neurodegeneration models.

Although a role for PSMC1 in Parkinson's disease pathogenesis has been suggested, current evidence suggests this association is unlikely , highlighting the importance of rigorous experimental design when investigating PSMC1 in specific neurodegenerative conditions.

How can PSMC1 antibodies be employed in cancer research, particularly relating to lung adenocarcinoma?

Recent research indicates that the PSMC family, including PSMC1, shows elevated expression in lung adenocarcinoma (LUAD) compared to normal lung tissues . This finding suggests potential roles for PSMC1 in cancer biology that warrant further investigation using PSMC1 antibodies:

• Expression profiling: PSMC1 antibodies can be used for immunohistochemical analysis of tissue microarrays to quantify PSMC1 expression across cancer stages and correlate with clinical outcomes.

• Prognostic marker investigation: Given that all PSMC genes show higher expression in LUAD tissues than in normal lung tissues , researchers should validate PSMC1 as a potential prognostic marker through studies correlating expression levels with patient survival data.

• Therapeutic target assessment: Researchers can use PSMC1 antibodies in drug development workflows to:

  • Screen compounds that modulate PSMC1 expression or function

  • Validate target engagement in drug-treated samples

  • Monitor changes in PSMC1 expression following experimental therapeutics

• Combination studies: Design experiments that examine PSMC1 in relation to other PSMC family members, particularly PSMC4, which shows the highest differential expression in LUAD (log2FC: 0.80) .

• Mechanism exploration: Use PSMC1 antibodies in combination with cell biology approaches to elucidate how increased PSMC1 expression contributes to cancer cell survival, proliferation, or treatment resistance.

What are the critical considerations for validating PSMC1 antibody specificity in experimental systems?

Antibody validation is essential for ensuring reliable and reproducible results. For PSMC1 antibodies, consider these validation approaches:

• Western blot validation: Confirm that the antibody detects a protein of the expected molecular weight (approximately 49 kDa for PSMC1). Look for single bands rather than multiple bands that might indicate cross-reactivity with related proteins.

• Knockdown/knockout controls: Use siRNA knockdown or CRISPR-Cas9 knockout of PSMC1 to verify antibody specificity. The signal should be significantly reduced or eliminated in these samples compared to controls.

• Peptide competition assays: Pre-incubate the antibody with the immunizing peptide (e.g., the synthetic peptide from amino acids 408-436 for ABIN1881692 ) before application to samples. This should block specific binding and eliminate true positive signals.

• Cross-reactivity assessment: If working with non-human samples, validate the antibody in your specific model organism. While many PSMC1 antibodies show cross-reactivity with mouse and rat samples , performance can vary across species.

• Recombinant protein standards: Use purified recombinant PSMC1 protein as a positive control in assays like western blotting or ELISA to establish detection limits and linear range.

• Multiple antibody comparison: When possible, use multiple antibodies targeting different epitopes of PSMC1. Concordant results provide stronger evidence of specificity.

How do different fixation and antigen retrieval methods affect PSMC1 antibody performance in immunohistochemistry?

The performance of PSMC1 antibodies in immunohistochemistry can be significantly affected by tissue processing methods:

• Fixation optimization: Compare paraformaldehyde, formalin, and alcohol-based fixatives to determine which best preserves PSMC1 antigenicity while maintaining tissue morphology. For the available PSMC1 antibodies, paraffin-embedded section protocols have been validated , suggesting compatibility with formalin fixation.

• Antigen retrieval methods: Test both heat-induced epitope retrieval (HIER) and enzymatic antigen retrieval:

  • For HIER, compare citrate buffer (pH 6.0) versus EDTA buffer (pH 9.0)

  • For enzymatic retrieval, evaluate proteinase K or trypsin digestion

  Document the optimal conditions for each PSMC1 antibody, as epitope accessibility may vary based on the target region (e.g., C-terminal epitopes may require different retrieval methods than internal epitopes).

• Incubation parameters: Systematically test antibody concentration, incubation time, and temperature to establish optimal signal-to-noise ratios. Start with manufacturer recommendations (if available) and adjust based on empirical results.

• Detection systems: Compare enzymatic (e.g., HRP-DAB) versus fluorescent detection systems to determine which provides optimal sensitivity and specificity for your specific research question.

• Special considerations for brain tissue: If studying neurodegenerative diseases, specialized fixation and staining protocols may be required due to the blood-brain barrier's effect on antibody penetration and the lipid-rich environment's impact on antigen accessibility.