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Western blotting

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Secondary Antibodies, Conjugates, and Kits
Technical Support for Secondary Antibodies, Conjugates, and Kits
Antibody Standard Validation
Explore antibody validation, including antibody and target, species validation, lot validation, application validation, storage and handling, citations, and quality control, in detail.
Antibody Enhanced-Validation Strategies
Antibody enhanced validation provides additional data to help ensure antibody specificity and performance and to address antibody reproducibility crisis.
Fluorescent Multiplex Detection using Antibody Atto Dye Conjugates
Immunoblotting (Western blot transfer) is a common technique in modern proteomics research.
Identification of Proteins in Lipofuscin Using Antibodies within the Human Protein Atlas
Identification of Proteins in Lipofuscin Using Antibodies within the Human Protein Atlas
Protein Visualization
Prior to probing a membrane using precious antibodies, it is helpful to visualize the transferred proteins on the blot to ensure complete transfer and even loading. This page describes possible causes and potential remedies for challenges encountered during protein visualization.
The possible causes and potential remedies for challenges encountered as a result of blocking, washing, antibody incubation, and detection/exposure of Western blots
Dot/Slot (Filtration) Blotting
The possible causes and potential remedies for challenges encountered dot/slot blotting, which uses vacuum filtration to transfer protein onto a microporous membrane.
Loading Controls for Western Blotting
Loading controls in western blotting application.
Using Conjugated and Secondary Antibodies
Using conjugated antibodies and secondary antibodies in the lab.
Methods Used to Detect Proteins and Nucleic Acids Bound to Membranes
Background and protocols describing the various methods used by molecular biologists to detect samples of protein or nucleic acids bound to membranes.
Antibody Basics
Explore the basics of working with antibodies including technical information on structure, classes, and normal immunoglobulin ranges.
Prestige Antibodies® Organelle Markers
Confocal microscopy analyzes protein subcellular localization in human cell lines.
Semi-Dry or Tank Electrotransfer Troubleshooting
Troubleshoot protein transfer issues in Western blotting by understanding causes and optimizing transfer conditions.
Cell Lysate Preparation
Cell lysis challenges and protein extraction variables for Western blot sensitivity and reproducibility.
Protein Staining Reagents Selection Guide
Sigma offers EZBlue™ and ProteoSilver™ reagents for protein visualization, suitable for proteomics and traditional PAGE formats.
Improving Reproducibility: Best Practices for Antibodies
In the midst of beeping lab timers, presentations and grant deadlines, it is easy to take for granted the quality of lab reagents.
Antibodies to Tubulins, including Centrosome-Associated ε-Tubulin
Microtubules of the eukaryotic cytoskeleton are composed of a heterodimer of α- and β-tubulin. In addition to α-and β-tubulin, several other tubulins have been identified, bringing the number of distinct tubulin classes to seven.
Alkaline Phosphatase
We offer a broad range of alkaline phosphatase enzymes and substrates that are optimized for conjugation to antibodies and other proteins for your specific application needs.
Colorimetric Detection of Western blot using 4-Chloro-1-Naphthol Solution Protocol
This solution is used as a substrate for detecting horseradish-peroxidase conjugates in Western blotting.
Anti-HA Troubleshooting
Anti-HA Troubleshooting
Western Blotting: Why Are Observed and Calculated Molecular Weights Different?
A discussion of factors affecting observed vs. calculated molecular weight in gel electrophoresis and western blotting.
How to Choose a Secondary Antibody
Review the key factors that should figure in your decision to choose a secondary antibody. Learn about species, subclass, isotype, label, and more.
An Introduction to Antibodies: Antigens, Epitopes and Antibodies
Antibodies combine with specific antigens to generate an exclusive antibody-antigen complex. Learn about the nature of this bond and its use as a molecular tag for research.
Prestige Antibodies® in Breast Cancer Research
Browse Human Protein Atlas; Prestige Antibodies®; Polyclonals and Monoclonals; Clinical Markers; Antibodies used in breast cancer research.
esiRNA Knockdown Efficiency Tested by Western Blotting
Quantitative and qualitative western blotting to validate knockdown by esiRNA.
Sample Prep & Gel Electrophoresis Troubleshooting
Identify causes and remedies for SDS-PAGE sample preparation challenges and optimize electrophoresis conditions.
Antibody Generation | Monoclonal vs Polyclonal Antibodies
Learn differences in monoclonal vs polyclonal antibodies including how antibodies are generated, clone numbers, and antibody formats.
Auto2D® Gel Electrophoresis Device
Auto2D® automates difficult 2D electrophoresis methods for protein localization with high reproducibility in under two hours.
Prestige Antibodies®: The Cell Atlas
Cell Atlas displays protein distribution with immunofluorescent staining in diverse cell lines, aiding spatial protein visualization.
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