Related Experiment Video

Updated: Feb 8, 2026

Preparation of Chloroplast Sub-compartments from Arabidopsis for the Analysis of Protein Localization by Immunoblotting or Proteomics
10:28

Preparation of Chloroplast Sub-compartments from Arabidopsis for the Analysis of Protein Localization by Immunoblotting or Proteomics

Published on: October 19, 2018

22.2K

Preparing Protein Solutions for Immunoblotting

Larisa Litovchick

    Cold Spring Harbor Protocols
    |July 4, 2018
    PubMed

    Most protein solutions can be mixed with sample buffer and analyzed directly by immunoblotting. Any protein extraction buffer or solvent that is compatible with gel electrophoresis can be used for preparing samples for immunoblotting. Examples of the protein solutions suitable for immunoblotting include purified proteins, detergent lysates of cells, samples prepared by subcellular fractionation, and protein eluates from chromatography columns. The protein of interest must be sufficiently abundant in the sample to allow for efficient detection, but the total amount of protein in the sample should not exceed the loading capacity of the gel. For this reason, protein antigens of very low abundance must be partially purified from the protein mixtures containing high concentrations of the contaminating proteins. In this protocol, solutions containing the protein antigens of interest are mixed with SDS-PAGE sample buffer and boiled to prepare samples for protein gel electrophoresis.

    More Related Videos

    Optimized Analysis of Proteins from Xenopus Oocytes and Embryos by Immunoblotting09:32

    Optimized Analysis of Proteins from Xenopus Oocytes and Embryos by Immunoblotting

    Published on: September 19, 2025

    596
    Quantitative Immunoblotting of Cell Lines as a Standard to Validate Immunofluorescence for Quantifying Biomarker Proteins in Routine Tissue Samples09:58

    Quantitative Immunoblotting of Cell Lines as a Standard to Validate Immunofluorescence for Quantifying Biomarker Proteins in Routine Tissue Samples

    Published on: January 7, 2019

    9.3K

    Related Concept Videos

    Ideal Solutions02:24

    Ideal Solutions

    According to Raoult’s law, the partial vapor pressure of a solvent in a solution is equal or identical to the vapor pressure of the pure solvent multiplied by its mole fraction in the solution. However, Raoult's Law is only valid for ideal solutions. For a solution to be ideal, the solvent-solute interaction must be just as strong as a solvent-solvent or solute-solute interaction. This suggests that both the solute and the solvent would use the same amount of energy to escape to the...
    22.6K
    General Properties of Solutions02:12

    General Properties of Solutions

    Many common substances around us exist as a solution, such as ocean water, air, and gasoline. All solutions are mixtures of substances that are composed of varying amounts of two or more types of atoms or molecules. A mixture with a non-uniform composition is a heterogeneous mixture, whereas a mixture with a uniform composition is a homogeneous mixture. The components that make the homogeneous mixture are evenly spread out and thoroughly mixed. 
    36.0K
    Solution Formation02:16

    Solution Formation

    There is no one solvent that can dissolve every type of solute. Some substances that readily dissolve in a certain solvent might be insoluble in a different solvent. A simple way to predict which substances dissolve in which solvent is the phrase "like dissolves like". This means that polar substances, such as salt and sugar, dissolve in a polar substance like water. In contrast, non-polar substances are more soluble in non-polar solvents such as carbon tetrachloride.
    This selective...
    37.9K
    Enthalpy of Solution02:39

    Enthalpy of Solution

    There are two criteria that favor, but do not guarantee, the spontaneous formation of a solution:
    31.1K
    Solution Concentration and Dilution02:59

    Solution Concentration and Dilution

    The relative amount of a given solution component is known as its concentration. Often, though not always, a solution contains one component with a concentration that is significantly greater than that of all other components. This component is called the solvent and may be viewed as the medium in which the other components are dispersed or dissolved. Solutions in which water is the solvent are, of course, very common on our planet. A solution in which water is the solvent is called an aqueous...
    134.7K
    Weak Base Solutions03:21

    Weak Base Solutions

    Some compounds produce hydroxide ions when dissolved by chemically reacting with water molecules. In all cases, these compounds react only partially and so are classified as weak bases. These types of compounds are also abundant in nature and important commodities in various technologies. For example, global production of the weak base ammonia is typically well over 100 metric tons annually, being widely used as an agricultural fertilizer, a raw material for chemical synthesis of other...
    25.3K