PRKCA Antibody, HRP conjugated
Description
Introduction
The PRKCA Antibody, HRP conjugated is a specialized immunological reagent designed for detecting protein kinase C alpha (PRKCA) in biological samples. PRKCA is a 77 kDa serine/threonine kinase involved in cellular signaling pathways, including cell proliferation, apoptosis, and metastasis . HRP (horseradish peroxidase) conjugation enhances the antibody’s utility in immunoassays by enabling enzymatic detection via chromogenic substrates like diaminobenzidine (DAB) or TMB .
Conjugation Methodology
HRP conjugation to PRKCA antibodies follows protocols like Abcam’s Lightning-Link® system . Key steps include:
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Buffer preparation: Antibody buffers must avoid components like BSA, gelatin, or Tris, which interfere with conjugation .
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Modifier addition: 1 µL of modifier per 10 µL antibody ensures efficient HRP coupling .
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Incubation and quenching: 3-hour incubation at room temperature, followed by reaction termination .
Prognostic Role in Lung Adenocarcinoma
High PRKCA protein expression correlates with poor prognosis in lung adenocarcinoma (LADC). A study of 123 LADC patients found co-expression of PRKCA and hepatoma-derived growth factor (HDGF) linked to shorter survival . Kaplan-Meier analysis revealed a hazard ratio of 2.45 (95% CI: 1.32–4.56) for high PRKCA/HDGF expression .
Mechanistic Insights
PRKCA regulates cellular processes like cell adhesion and survival . In LADC cells, PRKCA expression is upregulated by HDGF-mediated inhibition of miR-296-3p .
Applications in Immunological Techniques
Stability and Performance Optimization
HRP-conjugated antibodies degrade over time, exacerbated by temperature and dilution . Abcam’s LifeXtend™ stabilizer mitigates this by protecting the conjugate from environmental stressors .
