Western blotting

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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.

Anti-HA Troubleshooting

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.

Secondary Antibodies, Conjugates, and Kits

Technical Support for Secondary Antibodies, Conjugates, and Kits

Loading Controls for Western Blotting

Loading controls in western blotting application.

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.

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.

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.

Antibody Enhanced-Validation Strategies

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

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.

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.

Sample Prep & Gel Electrophoresis Troubleshooting

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

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.

Antibody Generation | Monoclonal vs Polyclonal Antibodies

Learn differences in monoclonal vs polyclonal antibodies including how antibodies are generated, clone numbers, and antibody formats.

Studying Hallmarks of Cancer With Recombinant Antibodies

Learn about the hallmarks of cancer and find reliable ZooMAb® recombinant antibodies to study them, conveniently organized by cancer hallmark.

Auto2D^® Gel Electrophoresis Device

Auto2D® automates difficult 2D electrophoresis methods for protein localization with high reproducibility in under two hours.

Immunodetection

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

Fluorescent Multiplex Detection using Antibody Atto Dye Conjugates

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

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.

Using Conjugated and Secondary Antibodies

Using conjugated antibodies and secondary antibodies in the lab.

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.

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

Prestige Antibodies® in Breast Cancer Research

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

Prestige Antibodies^® Organelle Markers

Confocal microscopy analyzes protein subcellular localization in human cell lines.

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.

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