Antibody-free Method
Antibody-free Method - Related Videos
10:13
Method for Whole Mount Antibody Staining in Chick
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Cited by 3 •
2009
This video demonstrates whole mount immunohistochemistry, a method by which the spatial and temporal expression pattern of an antigen can be visualized in young chick embryos. This method was originally introduced by Jane Dodd and Tom Jessell.
03:38
A Method to Produce Recombinant Antibodies Against Metallopeptidase
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2025
This video demonstrates a procedure for the production of recombinant antibodies against metallopeptidase. Recombinant plasmids carrying genes for the antibody, green fluorescent protein, and antibiotic resistance are transfected into epithelial cells with lipofectamine. The transfected cells are selected and cultivated to generate recombinant antibodies.
05:31
A Multiplex Immunostaining Method Using Primary Antibodies from the Same Host Species
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2025
This video demonstrates an assay to perform multiplex immunostaining using primary antibodies from the same host species. In this method, an adrenal tissue section undergoes a two-step immunostaining process. Initially, the section is stained with antibodies specific for markers in the cortex, followed by stripping the bound antibody and restaining with antibodies specific for markers in the medulla.
06:15
An In Vivo Immunofluorescence Localization Method to Study Antibody Biodistribution
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2025
This video demonstrates an in vivo immunofluorescence localization (IVIL) technique to study the biodistribution of tumor-targeting antibodies in a murine model of breast cancer. Upon the intravenous injection of a tumor-specific antibody in the mouse, the tumor is surgically isolated, and the localization of the antibody within the tumor tissue is confirmed via ex vivo immunostaining.
05:54
An In Vitro Method to Determine the Anti-Apoptotic Activity of Antibodies against TNFα
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2025
This video demonstrates a technique for assessing the efficacy of antibodies against tumor necrosis factor-alpha (TNFα), an inflammatory cytokine, through an in vitro bioanalytical approach. TNFα, when introduced to stressed cells, triggers apoptosis. However, binding to the antibodies confines TNFα, averting apoptosis. The degree of apoptosis is quantified using a bioluminescence assay.
FAQs
They distinguish molecules by measurable properties such as molecular mass, charge, amino acid sequence, chemical reactivity, or chromatographic behavior. Combining several of these characteristics increases the ability to separate and recognize related analytes within a complex sample. This principle is especially useful when different molecules have similar biological functions but differ in measurable chemical or physical features.
Enzymatic digestion can convert proteins into peptides that are more suitable for analysis. Liquid chromatography then separates components according to their chromatographic behavior, while mass spectrometry measures molecular mass and helps distinguish the resulting analytes. Used together, these steps provide complementary information for identifying or quantifying proteins and peptides without depending on antibody-antigen recognition.
An antibody-free method can be advantageous when suitable antibodies are unavailable, show variable performance, or lack sufficient selectivity. It also avoids dependence on custom antibody development, which can limit work on novel targets. By using direct molecular characteristics instead, the approach offers greater assay flexibility for biochemical measurements and target selection.
A supported workflow may begin with enzymatic digestion when protein-to-peptide conversion is appropriate, followed by liquid-chromatographic separation and mass-spectrometric measurement. The resulting data can be interpreted using molecular mass, sequence-related information, and chromatographic behavior to identify or quantify analytes. The exact combination depends on whether the goal is protein characterization, peptide analysis, or quantification.
These approaches support protein characterization, peptide analysis, post-translational modification studies, and quantitative proteomics. Their measurements can help researchers examine molecular composition and changes in modified or quantified species, including targets for which antibody development is impractical. This broad utility makes them relevant across biochemical investigations involving proteins and their derived peptides.
They are particularly useful when a target lacks an available antibody, when antibody selectivity is poor, or when antibody performance varies between measurements. Because the analysis can rely on molecular mass, sequence, charge, reactivity, or chromatographic behavior, researchers can investigate novel targets without first developing a custom recognition reagent. This expands experimental flexibility in biochemical research.