UQCRC2 Antibody

What is UQCRC2 and what cellular functions does it perform?

UQCRC2 functions as a core component of the ubiquinol-cytochrome c oxidoreductase, a multisubunit transmembrane complex that constitutes part of the mitochondrial electron transport chain driving oxidative phosphorylation. This protein associates with cytochrome b and cytochrome c1, collaborating in the electron transport chain to ensure efficient energy conversion processes within mitochondria. UQCRC2 is homologous to the alpha-MPP subunit of the mitochondrial processing peptidase and appears to retain some processing properties. Additionally, it likely plays a role in the in situ processing of UQCRFS1 into the mature Rieske protein when incorporated into complex III .

What experimental applications are UQCRC2 antibodies validated for?

Commercial UQCRC2 antibodies have been validated for multiple research applications with specific dilution ranges optimized for each technique:

It's important to note that antibody performance can vary between specific experimental systems, so researchers should perform optimization steps for their particular application and sample type .

What species reactivity do commercial UQCRC2 antibodies demonstrate?

The commercially available UQCRC2 antibodies show consistent reactivity across multiple mammalian species:

This cross-species reactivity makes these antibodies versatile tools for researchers working with various model organisms. The demonstrated reactivity with zebrafish samples also provides options for developmental biology studies .

How should UQCRC2 antibodies be stored for optimal stability and performance?

For maximum stability and retention of immunoreactivity, UQCRC2 antibodies should be stored according to manufacturer specifications:

• Store at -20°C in aliquots to minimize freeze-thaw cycles

• Stable for one year after shipment when properly stored

• Storage buffer typically consists of PBS with 0.02% sodium azide and 50% glycerol at pH 7.3

• For certain formulations, aliquoting may be unnecessary for -20°C storage

• Some preparations (e.g., 20μL sizes) may contain 0.1% BSA as a stabilizer

Proper storage conditions are critical for maintaining antibody performance across multiple experimental applications. Researchers should avoid repeated freeze-thaw cycles as they can lead to protein denaturation and diminished antibody activity .

What antigen retrieval methods are optimal for UQCRC2 detection in fixed tissues?

For optimal immunohistochemical detection of UQCRC2 in formalin-fixed paraffin-embedded (FFPE) tissues, specific antigen retrieval methods have been validated:

• Primary recommendation: Heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0

• Alternative method: Citrate buffer pH 6.0 may also be effective

• Protocol specifics: Perform heat-mediated antigen retrieval before commencing with IHC staining protocol

• Validated dilution range: 1:150-1:1200 depending on tissue type and detection system

These conditions have been successfully applied for detecting UQCRC2 in various human tissues including liver, colon, and colon cancer samples. The choice between Tris/EDTA and citrate buffer may depend on specific tissue characteristics and should be optimized accordingly .

How can researchers validate UQCRC2 antibody specificity using cellular controls?

Rigorous validation of antibody specificity is essential for reliable research outcomes. For UQCRC2 antibodies, multiple approaches can confirm specificity:

• Knockout/knockdown validation: Multiple publications (at least 2 cited) demonstrate successful validation using KD/KO approaches

• Western blot profile: Consistent detection of a single band at approximately 48 kDa (matching predicted molecular weight)

• Immunoprecipitation validation: Perform IP with UQCRC2 antibody followed by Western blot detection

• Negative controls: Include isotype control antibodies (e.g., rabbit monoclonal IgG) in parallel experiments

• Cross-validation: Compare staining patterns across multiple antibody clones targeting different UQCRC2 epitopes

For IP-based validation, researchers have successfully used UQCRC2 antibodies at 1:50 dilution for immunoprecipitation from 1mg of whole cell lysate (HepG2, HEK-293), followed by Western blot detection at 1:1000 dilution .

What evidence supports the involvement of UQCRC2 in cancer mechanisms?

Emerging research indicates significant connections between UQCRC2 and cancer pathobiology:

• Expression alterations: UQCRC2 expression levels are reported to be lower in gastric cancer (GC) tissues

• microRNA regulation: UQCRC2 appears to be regulated by miR-370, establishing the miR-370/UQCRC2 axis

• Functional impact: This regulatory axis may facilitate tumorigenesis by modulating metabolic processes

• Research applications: UQCRC2 antibodies serve as valuable tools for investigating these cancer-related mechanisms

• Tissue validation: Antibodies have been successfully applied to human colon cancer tissue versus normal colon

These findings suggest that UQCRC2 dysregulation may contribute to metabolic reprogramming in cancer cells, highlighting the importance of mitochondrial function in cancer development and progression .

What protocols are recommended for co-immunoprecipitation experiments using UQCRC2 antibodies?

For effective co-immunoprecipitation of UQCRC2 and its interacting partners:

• Sample preparation:

  • Prepare whole cell lysates from appropriate cells (validated in HepG2 and HEK-293 cells)

  • Use approximately 1mg of total protein for each IP reaction

• Immunoprecipitation:

  • Use UQCRC2 antibody at 1:50 dilution (approximately 0.5-4.0 μg antibody per reaction)

  • Incubate with protein lysate overnight at 4°C with gentle rotation

  • Capture antibody-protein complexes using appropriate beads (Protein A/G)

• Western blot detection:

  • Run IP samples alongside input control (10μg whole cell lysate)

  • Include negative control using isotype-matched irrelevant antibody

  • Detect using UQCRC2 antibody at 1:1000 dilution

  • Use secondary antibody specific to non-reduced form of IgG at 1:1500 dilution

This approach has been successfully validated for detecting UQCRC2 interactions and can be adapted to investigate its association with other mitochondrial complex III components .

How can researchers troubleshoot weak or inconsistent UQCRC2 signal in Western blot experiments?

When encountering suboptimal Western blot results with UQCRC2 antibodies, consider these systematic troubleshooting approaches:

• Sample preparation optimization:

  • UQCRC2 is reliably detected in various tissues including brain, heart, liver, and kidney

  • For cellular samples, HepG2, HEK-293, and Jurkat cells show strong expression

  • Ensure complete lysis of mitochondria using appropriate detergents

• Protocol adjustments:

  • Blocking: Use 5% non-fat dry milk in TBST as validated blocking agent

  • Primary antibody: Test dilution range between 1:2000-1:12000

  • Exposure time: Short exposures (3-5 seconds) are typically sufficient

  • Transfer conditions: Optimize for high molecular weight proteins (~48 kDa)

• Positive controls: Include validated positive controls such as:

  • Human: fetal brain, fetal heart, HepG2, HEK-293, or Jurkat cell lysates

  • Mouse/Rat: heart, brain, or kidney tissue lysates

• Detection system: Anti-Rabbit IgG HRP secondary antibody at 1:10000-1:50000 dilution has been validated for optimal detection .

What strategies improve signal-to-noise ratio in UQCRC2 immunofluorescence studies?

For optimal immunofluorescence detection of UQCRC2 in cellular samples:

• Fixation and permeabilization:

  • 100% methanol fixation followed by 0.1% Triton X-100 permeabilization has been validated

  • This approach preserves mitochondrial morphology while allowing antibody access

• Antibody conditions:

  • Primary antibody: Use at 1:50-1:500 dilution (validated at 1:300)

  • Secondary antibody: Alexa Fluor conjugated anti-rabbit IgG at 1:1000 dilution

• Controls and counterstaining:

  • Include DAPI nuclear counterstain

  • Consider co-staining with mitochondrial markers for colocalization studies

  • Include proper negative controls (secondary-only, isotype control)

• Imaging optimization:

  • Use confocal microscopy for optimal resolution of mitochondrial structures

  • Expected pattern: Cytoplasmic staining with mitochondrial distribution

  • Validated in multiple cell lines including HepG2 and Jurkat cells

Following these protocols should yield specific cytoplasmic staining corresponding to mitochondrial localization .

How can UQCRC2 antibodies be used to study mitochondrial dysfunction in disease models?

UQCRC2 antibodies provide valuable tools for investigating mitochondrial dysfunction across various disease contexts:

• Respiratory chain complex assessment:

  • UQCRC2 antibodies serve as reliable markers for complex III integrity

  • Can be used alongside markers for other respiratory complexes to assess specific defects

• Tissue-specific applications:

  • Differential expression analysis across tissues (validated in brain, heart, liver, kidney)

  • IHC applications in normal versus pathological tissues (e.g., colon cancer)

• Subcellular fractionation studies:

  • Mitochondrial isolation followed by Western blot analysis

  • Assessment of complex assembly through native gel electrophoresis

• Flow cytometric analysis:

  • Intracellular staining using paraformaldehyde fixation (validated at 1:150 dilution)

  • Allows quantitative assessment of UQCRC2 levels in cell populations

These approaches enable researchers to quantitatively assess mitochondrial complex III alterations in various disease models and experimental conditions .

How does miRNA regulation of UQCRC2 impact cellular metabolism and disease progression?

Emerging research has established important connections between miRNA regulation of UQCRC2 and disease pathophysiology:

• miR-370/UQCRC2 axis in cancer:

  • miR-370 appears to regulate UQCRC2 expression

  • This regulatory axis facilitates tumorigenesis in gastric cancer

  • UQCRC2 expression levels are lower in gastric cancer tissues

• Research approaches:

  • miRNA transfection studies coupled with UQCRC2 antibody detection

  • Assessment of mitochondrial function following miRNA manipulation

  • Correlation of UQCRC2 levels with clinical parameters in cancer studies

• Mechanistic implications:

  • Altered UQCRC2 expression may contribute to metabolic reprogramming in cancer

  • Mitochondrial dysfunction resulting from UQCRC2 dysregulation could affect oxidative phosphorylation

  • Changes in electron transport chain efficiency may influence cellular energy production

This emerging area represents an important intersection between mitochondrial biology, miRNA regulation, and cancer metabolism, highlighting the value of UQCRC2 antibodies in exploring these complex regulatory networks .