Loading Control

Loading Control

A loading control is a standard practice in many molecular biology and biochemistry experiments, particularly those involving techniques like Western blotting, gel electrophoresis, and quantitative PCR (qPCR). Its primary purpose is to ensure that equal amounts of protein or nucleic acid samples are loaded onto the gel or plate, thereby providing a reliable reference for comparing the expression levels of target molecules across different samples.

Here's how loading controls are used in different experimental techniques:

Western Blotting: In Western blotting, a loading control is typically a protein that is constitutively expressed at relatively constant levels across different experimental conditions or samples. Common loading control proteins include β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), tubulin, and histones. The loading control protein is detected alongside the target protein(s) of interest on the same membrane, and its signal intensity is used to normalize the signal from the target protein, accounting for variations in protein loading and transfer efficiency.

Gel Electrophoresis: In gel electrophoresis, loading controls are used to ensure that equal amounts of nucleic acids (DNA or RNA) are loaded onto the gel. For DNA gel electrophoresis, loading control markers of known sizes are often included alongside the sample lanes to confirm the correct migration of DNA fragments and assess the efficiency of electrophoresis. Similarly, for RNA gel electrophoresis, ribosomal RNA bands are often used as loading controls due to their consistent expression levels in most cell types.

Quantitative PCR (qPCR): In qPCR experiments, internal reference genes, also known as housekeeping genes, are used as loading controls to normalize gene expression levels across different samples. These reference genes, such as GAPDH, β-actin, or 18S rRNA, are stably expressed under various experimental conditions and serve as reliable indicators of the total RNA input. The expression levels of target genes are then normalized to the expression of the reference gene(s) to account for variations in RNA quantity and quality between samples.

 

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  • 12 Tag Protein Marker Lysate

    12 Tag Protein Marker Lysate | 175

    100

    NULL348.00
    12 Tag Protein Marker Lysate | 175 | Gentaur Species Reactivity: Multiple Epitope Tags Format: Lysate Product Type: Loading Control Source: E.coli
    100-175
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  • 12 Tag Protein Marker Lysate

    12 Tag Protein Marker Lysate | 45-LYS-Z

    100

    NULL348.00
    12 Tag Protein Marker Lysate | 45-LYS-Z | Gentaur Species Reactivity: Multiple Epitope Tags Format: Lysate Product Type: Loading Control Source: E.coli
    100-45-LYS-Z
    NULL348.00
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