UQCRFS1 Antibody

What is UQCRFS1 and what is its biological significance?

UQCRFS1 is a key subunit of the cytochrome bc1 complex (complex III) of the mitochondrial respiratory chain. The protein contains an iron-sulfur cluster that is essential for electron transport during oxidative phosphorylation. The UQCRFS1 gene encodes a 30 kDa protein with a transit peptide, though the observed molecular weight in experimental settings is typically around 23-25 kDa due to post-translational processing . Recent research has revealed its significance beyond basic cellular respiration, as it appears to be involved in the development of more aggressive phenotypes of breast cancer and has been linked to poor prognosis in ovarian cancer .

What are the alternative names and identifiers for UQCRFS1?

Researchers should be aware of the multiple nomenclature used in literature:

• RISP (Rieske Iron-Sulfur Protein)

• Complex III subunit 5

• Cytochrome b-c1 complex subunit 5

• Rieske iron-sulfur protein

• UQCRFS1 gene ID (NCBI): 7386

• UNIPROT ID: P47985

What species reactivity do commercial UQCRFS1 antibodies typically demonstrate?

Most commercially available UQCRFS1 antibodies show reactivity with human, mouse, and rat samples. Some antibodies have additionally been cited as reactive with pig samples . When selecting an antibody for your research, it's important to verify the specific species reactivity required for your experimental models, as effectiveness can vary between species despite listed cross-reactivity.

What are the validated applications for UQCRFS1 antibodies?

Current UQCRFS1 antibodies have been validated for several key applications:

Researchers should consider that Western Blot appears to be the most thoroughly validated application for studying UQCRFS1 across the literature .

What are the recommended dilutions for UQCRFS1 antibodies across different applications?

Optimal dilutions are sample-dependent, and researchers should perform titration experiments with their specific samples to determine optimal conditions . For novel tissue or cell types, a dilution series is strongly recommended to establish optimal signal-to-noise ratios.

What are the optimal storage conditions for UQCRFS1 antibodies?

Most UQCRFS1 antibodies should be stored at -20°C and are stable for one year after shipment. Notably, manufacturers specifically recommend against aliquoting certain antibodies . Storage buffers typically consist of PBS with 0.02% sodium azide and 50% glycerol at pH 7.3 . For 20 μl sized products, some may contain 0.1% BSA in the formulation . Always consult the manufacturer's product information for specific storage recommendations.

How does UQCRFS1 expression correlate with cancer progression and prognosis?

Research has demonstrated that UQCRFS1 is overexpressed in several cancer types, including gastric, breast, and ovarian cancers . In ovarian cancer specifically, high expression of UQCRFS1 has been associated with poor prognosis as determined by Kaplan-Meier analysis . Mechanistic studies revealed that UQCRFS1 expression correlates with cell cycle regulation, apoptosis, oxidative phosphorylation, and DNA damage responses. When UQCRFS1 was knocked down in ovarian cancer cell lines (A2780 and OVCAR8):

• Cell proliferation was reduced

• Cell cycle arrest occurred at the G1 phase

• Apoptosis rates increased

• ROS production increased

• DNA damage gene expression was elevated

• The AKT/mTOR pathway was inhibited

These findings suggest UQCRFS1 may represent a potential therapeutic target and prognostic biomarker in ovarian cancer research.

What are the challenges in detecting UQCRFS1 in different experimental systems?

When working with UQCRFS1 antibodies, researchers should be aware of several technical considerations:

• The calculated molecular weight (30 kDa) differs from the observed molecular weight (23-25 kDa) in most experimental systems due to post-translational modification and processing .

• For IHC applications, antigen retrieval conditions may significantly impact results. Some protocols suggest antigen retrieval with TE buffer pH 9.0, while others recommend citrate buffer pH 6.0 .

• When performing tissue-specific studies, validated positive controls include MCF-7 cells, mouse/rat heart tissue, and human stomach cancer tissue .

• Due to its role in mitochondrial function, proper subcellular fractionation techniques may be required for accurate localization studies.

What experimental controls should be included when studying UQCRFS1?

Rigorous experimental design requires appropriate controls:

What is the recommended Western blot protocol for UQCRFS1 detection?

For optimal Western blot results with UQCRFS1 antibodies:

• Sample preparation:

  • Extract proteins from cells/tissues using standard lysis buffers containing protease inhibitors

  • Include mitochondrial isolation steps if focusing specifically on mitochondrial fractions

• Gel electrophoresis:

  • Load 20-50 μg of total protein per lane

  • Use 10-12% SDS-PAGE gels for optimal resolution around the 23-25 kDa range

• Transfer and blocking:

  • Transfer to PVDF or nitrocellulose membranes

  • Block with 5% non-fat milk or BSA in TBST

• Antibody incubation:

  • Dilute primary antibody 1:1000-1:8000 in blocking buffer

  • Incubate overnight at 4°C

  • Wash thoroughly with TBST

  • Incubate with appropriate HRP-conjugated secondary antibody

• Detection:

  • Develop using enhanced chemiluminescence

  • Expected band size: 23-25 kDa

For complete protocol details, manufacturers provide product-specific protocols that can be downloaded .

What are the recommended protocols for immunoprecipitation of UQCRFS1?

For successful immunoprecipitation of UQCRFS1:

• Sample preparation:

  • Lyse cells/tissues in non-denaturing lysis buffer

  • Use 1.0-3.0 mg of total protein lysate

• Antibody binding:

  • Add 0.5-4.0 μg of UQCRFS1 antibody per sample

  • Alternatively, use dilution of 1:50

  • Incubate with rotation overnight at 4°C

• Immunoprecipitation:

  • Add protein A/G beads

  • Incubate with rotation for 2-4 hours at 4°C

  • Wash beads thoroughly with lysis buffer

  • Elute proteins with SDS sample buffer

• Analysis:

  • Perform Western blot as described earlier

  • Mouse heart tissue has been validated as a positive control for IP experiments

Complete IP protocols specific to UQCRFS1 antibodies are available from manufacturers .

What are the recommended immunohistochemistry protocols for UQCRFS1?

For successful IHC detection of UQCRFS1:

• Tissue preparation:

  • Fix tissues in 10% neutral buffered formalin

  • Embed in paraffin and section at 4-6 μm thickness

• Antigen retrieval:

  • Use TE buffer pH 9.0 (recommended)

  • Alternative: citrate buffer pH 6.0

  • Heat-induced epitope retrieval methods are typically effective

• Antibody incubation:

  • Dilute primary antibody 1:250-1:1000 or 1:20-1:200 in antibody diluent

  • Incubate overnight at 4°C or as recommended by manufacturer

  • Wash thoroughly with PBS or TBST

• Detection:

  • Use appropriate detection system (HRP/DAB, etc.)

  • Human stomach cancer tissue has been validated as a positive control

Detailed IHC protocols specific to UQCRFS1 antibodies are available from manufacturers .

How is UQCRFS1 being studied in cancer research?

UQCRFS1 is emerging as an important target in cancer research due to its differential expression and association with patient outcomes. Key research areas include:

• Expression profiling: Evaluating UQCRFS1 expression across different cancer types and correlating with clinical parameters.

• Prognostic biomarker development: Studies have shown UQCRFS1 expression correlates with poor prognosis in ovarian cancer, suggesting potential as a biomarker .

• Functional studies: Knockdown experiments have revealed roles in cell proliferation, cell cycle regulation, apoptosis, and ROS production .

• Pathway analysis: Research has demonstrated UQCRFS1's involvement in the AKT/mTOR pathway, suggesting potential therapeutic implications .

• Therapeutic targeting: Emerging research is exploring whether UQCRFS1 could be directly targeted or used as a biomarker for targeted therapy selection.

What are the emerging techniques for studying UQCRFS1 function?

Advanced methodologies being applied to UQCRFS1 research include:

• CRISPR/Cas9 gene editing: For creating knockout and knock-in cell lines to study function.

• Mitochondrial respiratory chain complex assembly analysis: To understand how UQCRFS1 contributes to mitochondrial function in normal and disease states.

• Proximity labeling approaches: To identify novel protein interactions of UQCRFS1 within the mitochondrial network.

• Patient-derived xenografts: For testing therapeutic approaches targeting UQCRFS1 in more physiologically relevant models.

• Multi-omics integration: Combining proteomics, transcriptomics, and metabolomics to understand the broader impact of UQCRFS1 dysregulation.

How does UQCRFS1 interact with other components of the respiratory chain?

UQCRFS1 plays a critical role in the electron transport chain as part of Complex III (cytochrome bc1 complex). Research has shown that:

• It contains the Rieske iron-sulfur cluster that is essential for electron transfer from ubiquinol to cytochrome c .

• Its proper function is critical for maintaining mitochondrial bioenergetics and preventing excessive ROS production.

• UQCRFS1 coordination with other respiratory complexes is essential for supercomplex assembly and optimal mitochondrial function, as demonstrated in cardioprotection studies .

• Research has linked UQCRFS1 with the NRF1-PGC1α complex in breast cancer, suggesting a role in mitochondrial biogenesis regulation .

Further research is needed to fully elucidate the interactome of UQCRFS1 and its role in respiratory chain assembly and function in both normal and pathological conditions.