Western blotting

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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.

Anti-HA Troubleshooting

Fluorescent Multiplex Detection using Antibody Atto Dye Conjugates

Immunoblotting (Western blot transfer) is a common technique in modern proteomics research.

Immunodetection

The possible causes and potential remedies for challenges encountered as a result of blocking, washing, antibody incubation, and detection/exposure of Western blots

Identification of Proteins in Lipofuscin Using Antibodies within the Human Protein Atlas

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.

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.

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.

Antibody Enhanced-Validation Strategies

Antibody enhanced validation provides additional data to help ensure antibody specificity and performance and to address antibody reproducibility crisis.

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.

Antibody Basics

Explore the basics of working with antibodies including technical information on structure, classes, and normal immunoglobulin ranges.

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.

esiRNA Knockdown Efficiency Tested by Western Blotting

Quantitative and qualitative western blotting to validate knockdown by esiRNA.

Prestige Antibodies®: The Cell Atlas

Cell Atlas displays protein distribution with immunofluorescent staining in diverse cell lines, aiding spatial protein visualization.

Protein Staining Reagents Selection Guide

Sigma offers EZBlue™ and ProteoSilver™ reagents for protein visualization, suitable for proteomics and traditional PAGE formats.

Water for Protein Electrophoresis & Western Blotting

Explore the impact of water quality on protein electrophoresis and Western blotting. Learn about the role of ultrapure water and its effect on chemiluminescent signals in Western blotting.

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.

Antibodies in Practice: Conjugated and Secondary Antibody Use

Using conjugated antibodies and secondary antibodies in the lab.

Loading Controls for Western Blotting

Loading controls in western blotting application.

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.

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.

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.

Prestige Antibodies® in Breast Cancer Research

Browse Human Protein Atlas; Prestige Antibodies®; Polyclonals and Monoclonals; Clinical Markers; Antibodies used in breast cancer research.

Sample Prep & Gel Electrophoresis Troubleshooting

Identify causes and remedies for SDS-PAGE sample preparation challenges and optimize electrophoresis conditions.

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.

Western Blot Analysis of Immunoprecipitation (IP-Western)

The possible causes and potential remedies for challenges encountered in the immunoprecipitation-Western blot technique, which consists of cell lysis, formation of the antibody-antigen (immune) complex, precipitation of the immune complexes, and analysis by Western blotting.

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