Using multiple primary antibodies in complex immunoassays presents challenges, such as cross-reactivity and limited multiplexing options. One strategy involves selecting primary antibodies from different host species, like guinea pig and rabbit, to enable distinct recognition by secondary antibodies. An alternative approach includes the incorporation of primary antibodies labeled with distinct fluorescent dyes, which eliminates unwanted secondary antibody cross-reactivity, streamlining your process and reducing potential issues.
Guinea pig primary antibodies offer a versatile approach for complex immunodetection scenarios, particularly in colocalization, multiplex immunofluorescence and complex immunoprecipitation studies. Combining antibodies from different hosts, including guinea pig, rabbit, and mouse, minimizes cross-reactivity to ensure accurate targeting of multiple proteins within a single sample.
At Alomone Labs, we offer a variety of guinea pig primary antibodies, along with rabbit and mouse antibodies. This allows you to simultaneously target multiple antigens using primary antibodies from different hosts.
Incorporating fluorophore-conjugated primary antibodies into immunoassays is an additional valuable approach for multiplex immunostaining and immunodetection. Fluorophore-conjugated antibodies can be paired with an unconjugated primary antibody (regardless of the host species), or with an antibody conjugated to a different fluorophore, for experiments associated with immunolabeling of multiple antigens. These antibodies simplify the labeling process and save both reagents and time. Learn more about Alomone’s conjugated antibodies and customized solutions.
Cat # | APR-095-F |
Host | Rabbit |
Clonality | Polyclonal |
Cat # | APR-095-F |
Host | Rabbit |
Clonality | Polyclonal |
Cat # | APR-095-F |
Host | Rabbit |
Clonality | Polyclonal |
Microglia, the resident macrophages of the central nervous system (CNS), make up approximately 10% of the CNS cell population and play a critical role in maintaining homeostasis, responding to injury and inflammation, and clearing cellular debris.
Dysregulation of microglia function has been implicated in a range of neurological disorders, including Alzheimer’s disease and multiple sclerosis. Given their vital function in brain health and disease, there is a growing interest in developing tools and techniques for identifying and studying microglia.
Microglia markers are specific proteins or molecules that are expressed by microglia and that are commonly used to identify and study these cells, to distinguish microglia from other cells, and to provide insights into dysfunctional states. The use of microglia markers can also help identify changes in microglial function that occur during injury or disease.
Microglia-specific markers include surface markers, intracellular markers, and functional markers. Surface markers are proteins found on the outer membrane of microglial cells and play a crucial role in identifying and characterizing cell types, activation and functional states, and disease-related implications. Intracellular markers are located inside microglial cells and are used to investigate specific cellular processes, signaling pathways, and activation states. Functional markers are associated with specific microglial functions, including inflammatory response, neuroprotection, tissue repair, and disease-associated changes. Microglia-specific markers serve as valuable tools for research, diagnostics, and potential therapeutic interventions targeting microglial activity in neurological conditions.
A widely used technique to analyze microglia markers is immunofluorescence (IF), which allows for the visualization and localization of specific molecules within cells or tissues. IF uses fluorescently labeled antibodies to detect and target specific proteins or antigens of interest. The obtained fluorescence images can be analyzed and quantified using image analysis software. Colocalization analysis can determine the degree of overlap between different microglia-specific markers, indicating potential interactions or colocalization within cells or tissues. Fluorescence intensity measurements can provide insights into relative protein expression levels and their subcellular localization patterns.
Microglia markers play a useful role in investigating various neurological diseases. Microglia play a crucial role in the pathogenesis of Alzheimer’s disease, which is characterized by the accumulation of amyloid beta plaques and neurofibrillary tangles in the brain. Under normal conditions, microglia can phagocytose and clear amyloid beta plaques. In Alzheimer’s disease, microglia often exhibit an impaired ability to efficiently remove these toxic protein aggregates. The dysfunction of microglia in Alzheimer’s disease is believed to be a result of the chronic inflammatory environment and dysregulated immune response, which can exacerbate neurodegeneration.
In Parkinson’s disease, a neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra of the brain, microglia impairment decreases the clearance capacity of Lewy bodies, misfolded alpha-synuclein aggregates which are the hallmark of Parkinson’s disease. Chronic neuroinflammation mediated by microglia is thought to contribute to the progression of neurodegeneration in Parkinson’s disease.
Microglia respond to CNS injuries, such as traumatic brain injury (TBI) and spinal cord injury. Upon injury, microglia become activated and undergo morphological changes, migrating to the site of injury and releasing various factors, such as cytokines, chemokines, and reactive oxygen species, which can contribute to secondary damage and neuroinflammation after CNS injuries. Microglia also have beneficial functions in CNS injuries, such as promoting tissue repair, clearance of cellular debris, and releasing neurotrophic factors that support neuronal survival and regeneration. Modulating microglial activation and inflammatory responses is an area of active research for potential therapeutic interventions for CNS injuries.